104 results on '"Zaehres, H."'
Search Results
2. Generation of two human iPSC lines (HIMRi002-A and HIMRi003-A) derived from Caveolinopathy patients with rippling muscle disease
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Boeing, A., primary, Mavrommatis, L., additional, Daya, N.M., additional, Zhuge, H., additional, Volke, L., additional, Kocabas, A., additional, Kneifel, M., additional, Athamneh, M., additional, Krause, K., additional, Südkamp, N., additional, Döring, K., additional, Theiss, C., additional, Roos, A., additional, Zaehres, H., additional, Güttsches, A.K., additional, and Vorgerd, M., additional
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- 2023
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3. Generation of a human iPSC line (HIMRi001-A) from a patient with filaminopathy
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Daya, N.M., primary, Mavrommatis, L., additional, Zhuge, H., additional, Athamneh, M., additional, Roos, A., additional, Gläser, D., additional, Doering, K., additional, Zaehres, H., additional, Vorgerd, M., additional, and Güttsches, A.K., additional
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- 2023
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4. Generation of two hiPSCs lines of two patients carrying truncating mutations in the dimerization domain of filamin C
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Daya, N.M., Döring, K., Zhuge, H., Volke, L., Stab, V., Dietz, J., Athamneh, M., Roos, A., Zaehres, H., Güttsches, A.K., Mavrommatis, L., and Vorgerd, M.
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- 2024
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5. NANOG promotes cancer stem cell characteristics and prostate cancer resistance to androgen deprivation
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Jeter, C R, Liu, B, Liu, X, Chen, X, Liu, C, Calhoun-Davis, T, Repass, J, Zaehres, H, Shen, J J, and Tang, D G
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- 2011
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6. Transcription factor: induced pluripotency: KL043
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Zaehres, H, Kim, J B, Greber, B, Wu, G, Bravo, Arauzo M, Singhal, N, Cantz, T, Bleidissel, M, Fischedick, G, Zeuschner, D, Sebastiano, V, Gentile, L, Do, J T, Ko, K, Han, D W, Sterneckert, J, Ruau, D, Sasse, P, Meyer, J, Fleischmann, B K, Zenke, M, and Schöler, H R
- Published
- 2009
7. Influence of thermal annealing on the martensitic transitions in Ni–Ti shape memory alloys
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Somsen, Ch, Zähres, H, Kästner, J, Wassermann, E.F, Kakeshita, T, and Saburi, T
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- 1999
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8. GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells
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Wal, Erik, Bergsma, Atze, Gestel, Tom, in 't Groen, Stijn, Zaehres, H, Arauzo-Bravo, MJ, Scholer, HR, van der Ploeg, Ans, Pijnappel, Pim, Wal, Erik, Bergsma, Atze, Gestel, Tom, in 't Groen, Stijn, Zaehres, H, Arauzo-Bravo, MJ, Scholer, HR, van der Ploeg, Ans, and Pijnappel, Pim
- Published
- 2017
9. Antisense oligonucleotides promote exon inclusion and correct the common c.-32-13T > G (IVS1) GAA splicing variant in iPS-derived skeletal muscle cells from Pompe patients
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Pijnappel, W., primary, van der Wal, E., additional, Bergsma, A., additional, van Gestel, T., additional, Pijnenburg, J., additional, In ‘t Groen, S., additional, Zaehres, H., additional, Araúzo-Bravo, M., additional, Schöler, H., additional, and van der Ploeg, A., additional
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- 2017
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10. P.218 - Antisense oligonucleotides promote exon inclusion and correct the common c.-32-13T > G (IVS1) GAA splicing variant in iPS-derived skeletal muscle cells from Pompe patients
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Pijnappel, W., van der Wal, E., Bergsma, A., van Gestel, T., Pijnenburg, J., In ‘t Groen, S., Zaehres, H., Araúzo-Bravo, M., Schöler, H., and van der Ploeg, A.
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- 2017
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11. Erythroid differentiation of human induced pluripotent stem cells is independent of donor cell type of origin
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Dorn, I., primary, Klich, K., additional, Arauzo-Bravo, M. J., additional, Radstaak, M., additional, Santourlidis, S., additional, Ghanjati, F., additional, Radke, T. F., additional, Psathaki, O. E., additional, Hargus, G., additional, Kramer, J., additional, Einhaus, M., additional, Kim, J. B., additional, Kogler, G., additional, Wernet, P., additional, Scholer, H. R., additional, Schlenke, P., additional, and Zaehres, H., additional
- Published
- 2014
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12. A central role for TFIID in the pluripotent transcription circuitry
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Pijnappel, W.W.M.P., Esch, D., Baltissen, M.P.A., Wu, G., Mischerikow, N., Bergsma, A.J., Wal, E. van de, Han, D.W., Bruch, H., Moritz, S., Lijnzaad, P., Altelaar, A.F., Sameith, K., Zaehres, H., Heck, A.J.R. van, Holstege, F.C., Scholer, H.R., Timmers, H.T.M., Pijnappel, W.W.M.P., Esch, D., Baltissen, M.P.A., Wu, G., Mischerikow, N., Bergsma, A.J., Wal, E. van de, Han, D.W., Bruch, H., Moritz, S., Lijnzaad, P., Altelaar, A.F., Sameith, K., Zaehres, H., Heck, A.J.R. van, Holstege, F.C., Scholer, H.R., and Timmers, H.T.M.
- Abstract
Item does not contain fulltext, Embryonic stem (ES) cells are pluripotent and characterized by open chromatin and high transcription levels, achieved through auto-regulatory and feed-forward transcription factor loops. ES-cell identity is maintained by a core of factors including Oct4 (also known as Pou5f1), Sox2, Klf4, c-Myc (OSKM) and Nanog, and forced expression of the OSKM factors can reprogram somatic cells into induced pluripotent stem cells (iPSCs) resembling ES cells. These gene-specific factors for RNA-polymerase-II-mediated transcription recruit transcriptional cofactors and chromatin regulators that control access to and activity of the basal transcription machinery on gene promoters. How the basal transcription machinery is involved in setting and maintaining the pluripotent state is unclear. Here we show that knockdown of the transcription factor IID (TFIID) complex affects the pluripotent circuitry in mouse ES cells and inhibits reprogramming of fibroblasts. TFIID subunits and the OSKM factors form a feed-forward loop to induce and maintain a stable transcription state. Notably, transient expression of TFIID subunits greatly enhanced reprogramming. These results show that TFIID is critical for transcription-factor-mediated reprogramming. We anticipate that, by creating plasticity in gene expression programs, transcription complexes such as TFIID assist reprogramming into different cellular states.
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- 2013
13. A central role for TFIID in the pluripotent transcription circuitry
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Biomolecular Mass Spectrometry and Proteomics, Sub Biomol.Mass Spect. and Proteomics, Pijnappel, W.W.M., Esch, D., Baltissen, M.P.A., Wu, G., Mischerikow, N.|info:eu-repo/dai/nl/311489869, Bergsma, A., van der Wal, E., Han, D.W., vom Bruch, H., Moritz, S., Lijnzaad, P., Altelaar, A.F.M.|info:eu-repo/dai/nl/304833517, Sameith, K., Zaehres, H., Heck, A.J.R.|info:eu-repo/dai/nl/105189332, Holstege, F.C.P., Scholer, H.R., Timmers, H.T.M., Biomolecular Mass Spectrometry and Proteomics, Sub Biomol.Mass Spect. and Proteomics, Pijnappel, W.W.M., Esch, D., Baltissen, M.P.A., Wu, G., Mischerikow, N.|info:eu-repo/dai/nl/311489869, Bergsma, A., van der Wal, E., Han, D.W., vom Bruch, H., Moritz, S., Lijnzaad, P., Altelaar, A.F.M.|info:eu-repo/dai/nl/304833517, Sameith, K., Zaehres, H., Heck, A.J.R.|info:eu-repo/dai/nl/105189332, Holstege, F.C.P., Scholer, H.R., and Timmers, H.T.M.
- Published
- 2013
14. Magnetic and elastic properties of superparamagnetic CoMn-alloys
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Imbierwitz, F., Pelzl, J., Arnscheidt, E., Acet, M., Zähres, H., Bach, H., and Saunders, G.A.
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- 1999
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15. The magnetic states and magnetoelasticity in bcc invar-type Cr75Fe25−xMnx alloys
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Acet, M., Schneider, T., Zähres, H., Stamm, W., Wassermann, E.F., and Pepperhoff, W.
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- 1990
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16. The study of the thermal properties of γ-Mn from Mn-Cu alloys
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Acet, M., Zähres, H., Stamm, W., Wassermann, E.F., and Pepperhoff, W.
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- 1990
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17. Magnetovolume effects in Fe 80- xNi xCr 20 alloys
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Acet, M., Stamm, W., Zähres, H., and Wassermann, E.F.
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- 1987
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18. Coexisting antiferromagnetism and ferromagnetism in Fe-Ni Invar
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Zähres, H., Acet, M., Stam, W., and Wassermann, E.F.
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- 1988
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19. Magnetovolume effects in Fe80−xNixCr20 alloys
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Acet, M., Stamm, W., Wassermann, E.F., and Zähres, H.
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- 1987
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20. Thermal expansion of Fe50NixMn50−x
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Wassermann, E.F., Zähres, H., Acet, M., and Stamm, W.
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- 1987
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21. Nuclear and cytosolic fractions of SOX2 synergize as transcriptional and translational co-regulators of cell fate.
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Schaefer T, Mittal N, Wang H, Ataman M, Candido S, Lötscher J, Velychko S, Tintignac L, Bock T, Börsch A, Baßler J, Rao TN, Zmajkovic J, Roffeis S, Löliger J, Jacob F, Dumlin A, Schürch C, Schmidt A, Skoda RC, Wymann MP, Hess C, Schöler HR, Zaehres H, Hurt E, Zavolan M, and Lengerke C
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- Humans, Cell Differentiation, Animals, Protein Biosynthesis, Mice, SOXB1 Transcription Factors metabolism, SOXB1 Transcription Factors genetics, Cytosol metabolism, Cell Nucleus metabolism, Transcription, Genetic
- Abstract
Stemness and pluripotency are mediated by transcriptional master regulators that promote self-renewal and repress cell differentiation, among which is the high-mobility group (HMG) box transcription factor SOX2. Dysregulated SOX2 expression, by contrast, leads to transcriptional aberrations relevant to oncogenic transformation, cancer progression, metastasis, therapy resistance, and relapse. Here, we report a post-transcriptional mechanism by which the cytosolic pool of SOX2 contributes to these events in an unsuspected manner. Specifically, a low-complexity region within SOX2's C-terminal segment connects to the ribosome to modulate the expression of cognate downstream factors. Independent of nuclear structures or DNA, this C-terminal functionality alone changes metabolic properties and induces non-adhesive growth when expressed in the cytosol of SOX2 knockout cells. We thus propose a revised model of SOX2 action where nuclear and cytosolic fractions cooperate to impose cell fate decisions via both transcriptional and translational mechanisms., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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22. Generation of Skeletal Muscle Organoids from Human Pluripotent Stem Cells.
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Kindler U, Zaehres H, and Mavrommatis L
- Abstract
Various protocols have been proven effective in the directed differentiation of mouse and human pluripotent stem cells into skeletal muscles and used to study myogenesis. Current 2D myogenic differentiation protocols can mimic muscle development and its alteration under pathological conditions such as muscular dystrophies. 3D skeletal muscle differentiation approaches can, in addition, model the interaction between the various cell types within the developing organoid. Our protocol ensures the differentiation of human embryonic/induced pluripotent stem cells (hESC/hiPSC) into skeletal muscle organoids (SMO) via cells with paraxial mesoderm and neuromesodermal progenitors' identity and further production of organized structures of the neural plate margin and the dermomyotome. Continuous culturing omits neural lineage differentiation and promotes fetal myogenesis, including the maturation of fibroadipogenic progenitors and PAX7-positive myogenic progenitors. The PAX7 progenitors resemble the late fetal stages of human development and, based on single-cell transcriptomic profiling, cluster close to adult satellite cells of primary muscles. To overcome the limited availability of muscle biopsies from patients with muscular dystrophy during disease progression, we propose to use the SMO system, which delivers a stable population of skeletal muscle progenitors from patient-specific iPSCs to investigate human myogenesis in healthy and diseased conditions. Key features • Development of skeletal muscle organoid differentiation from human pluripotent stem cells, which recapitulates myogenesis. • Analysis of early embryonic and fetal myogenesis. • Provision of skeletal muscle progenitors for in vitro and in vivo analysis for up to 14 weeks of organoid culture. • In vitro myogenesis from patient-specific iPSCs allows to overcome the bottleneck of muscle biopsies of patients with pathological conditions., Competing Interests: Competing interestsThe authors declare that there are no conflicts of interest regarding the publication of this paper., (©Copyright : © 2024 The Authors; This is an open access article under the CC BY license.)
- Published
- 2024
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23. Osteopontin drives neuroinflammation and cell loss in MAPT-N279K frontotemporal dementia patient neurons.
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Al-Dalahmah O, Lam M, McInvale JJ, Qu W, Nguyen T, Mun JY, Kwon S, Ifediora N, Mahajan A, Humala N, Winters T, Angeles E, Jakubiak KA, Kühn R, Kim YA, De Rosa MC, Doege CA, Paryani F, Flowers X, Dovas A, Mela A, Lu H, DeTure MA, Vonsattel JP, Wszolek ZK, Dickson DW, Kuhlmann T, Zaehres H, Schöler HR, Sproul AA, Siegelin MD, De Jager PL, Goldman JE, Menon V, Canoll P, and Hargus G
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- Humans, Animals, Mice, Neuroinflammatory Diseases metabolism, Neuroinflammatory Diseases pathology, Microglia metabolism, Microglia pathology, Mutation genetics, Osteopontin metabolism, Osteopontin genetics, Frontotemporal Dementia genetics, Frontotemporal Dementia pathology, Frontotemporal Dementia metabolism, Neurons metabolism, Neurons pathology, tau Proteins metabolism
- Abstract
Frontotemporal dementia (FTD) is an incurable group of early-onset dementias that can be caused by the deposition of hyperphosphorylated tau in patient brains. However, the mechanisms leading to neurodegeneration remain largely unknown. Here, we combined single-cell analyses of FTD patient brains with a stem cell culture and transplantation model of FTD. We identified disease phenotypes in FTD neurons carrying the MAPT-N279K mutation, which were related to oxidative stress, oxidative phosphorylation, and neuroinflammation with an upregulation of the inflammation-associated protein osteopontin (OPN). Human FTD neurons survived less and elicited an increased microglial response after transplantation into the mouse forebrain, which we further characterized by single nucleus RNA sequencing of microdissected grafts. Notably, downregulation of OPN in engrafted FTD neurons resulted in improved engraftment and reduced microglial infiltration, indicating an immune-modulatory role of OPN in patient neurons, which may represent a potential therapeutic target in FTD., Competing Interests: Declaration of interests Z.K.W. serves as PI or co-PI on Biohaven Pharmaceuticals, Inc. (BHV4157-206), Vigil Neuroscience, Inc. (VGL101-01.002, VGL101-01.201), and ONO-2808-03 projects/grants. He serves as co-PI of the Mayo Clinic APDA Center for Advanced Research and as an external advisory board member for Vigil Neuroscience, Inc. and as a consultant for Eli Lilly & Company and for NovoGlia, Inc., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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24. Human skeletal muscle organoids model fetal myogenesis and sustain uncommitted PAX7 myogenic progenitors.
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Mavrommatis L, Jeong HW, Kindler U, Gomez-Giro G, Kienitz MC, Stehling M, Psathaki OE, Zeuschner D, Bixel MG, Han D, Morosan-Puopolo G, Gerovska D, Yang JH, Kim JB, Arauzo-Bravo MJ, Schwamborn JC, Hahn SA, Adams RH, Schöler HR, Vorgerd M, Brand-Saberi B, and Zaehres H
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- Humans, Cell Differentiation, Fetus metabolism, Muscle Development physiology, PAX7 Transcription Factor metabolism, Muscle, Skeletal metabolism, Satellite Cells, Skeletal Muscle physiology
- Abstract
In vitro culture systems that structurally model human myogenesis and promote PAX7
+ myogenic progenitor maturation have not been established. Here we report that human skeletal muscle organoids can be differentiated from induced pluripotent stem cell lines to contain paraxial mesoderm and neuromesodermal progenitors and develop into organized structures reassembling neural plate border and dermomyotome. Culture conditions instigate neural lineage arrest and promote fetal hypaxial myogenesis toward limb axial anatomical identity, with generation of sustainable uncommitted PAX7 myogenic progenitors and fibroadipogenic (PDGFRa+) progenitor populations equivalent to those from the second trimester of human gestation. Single-cell comparison to human fetal and adult myogenic progenitor /satellite cells reveals distinct molecular signatures for non-dividing myogenic progenitors in activated ( CD44High / CD98+ / MYOD1+ ) and dormant ( PAX7High / FBN1High / SPRY1High ) states. Our approach provides a robust 3D in vitro developmental system for investigating muscle tissue morphogenesis and homeostasis., Competing Interests: LM, HJ, UK, GG, MK, MS, OP, DZ, MB, DH, GM, DG, JK, MA, JS, SH, RA, HS, MV, BB, HZ No competing interests declared, JY is partially employed by Next & Bio Inc, (© 2023, Mavrommatis et al.)- Published
- 2023
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25. CRISPR/Cas9 Genome Editing in LGMD2A/R1 Patient-Derived Induced Pluripotent Stem and Skeletal Muscle Progenitor Cells.
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Mavrommatis L, Zaben A, Kindler U, Kienitz MC, Dietz J, Jeong HW, Böhme P, Brand-Saberi B, Vorgerd M, and Zaehres H
- Abstract
Large numbers of Calpain 3 (CAPN3) mutations cause recessive forms of limb-girdle muscular dystrophy (LGMD2A/LGMDR1) with selective atrophy of the proximal limb muscles. We have generated induced pluripotent stem cells (iPSC) from a patient with two mutations in exon 3 and exon 4 at the calpain 3 locus (W130C, 550delA). Two different strategies to rescue these mutations are devised: (i) on the level of LGMD2A-iPSC, we combined CRISPR/Cas9 genome targeting with a FACS and Tet transactivator-based biallelic selection strategy, which resulted in a new functional chimeric exon 3-4 without the two CAPN3 mutations. (ii) On the level of LGMD2A-iPSC-derived CD82+/Pax7+ myogenic progenitor cells, we demonstrate CRISPR/Cas9 mediated rescue of the highly prevalent exon 4 CAPN3 mutation. The first strategy specifically provides isogenic LGMD2A corrected iPSC for disease modelling, and the second strategy can be further elaborated for potential translational approaches., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2023 Lampros Mavrommatis et al.)
- Published
- 2023
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26. Interactive teaching enhances students' physiological arousal during online learning.
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Gellisch M, Morosan-Puopolo G, Wolf OT, Moser DA, Zaehres H, and Brand-Saberi B
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- Humans, Learning, Curriculum, Arousal, Teaching, Education, Distance methods, Students, Medical
- Abstract
The pure transfer of face-to-face teaching to a digital learning environment can be accompanied by a significant reduction in the physiological arousal of students, which in turn can be associated with passivity during the learning process, often linked to insufficient levels of concentration and engagement in the course work. Therefore, the aim of this study was to investigate whether students' psychobiological stress responses can be enhanced in the context of anatomical online learning and how increased physiological parameters correlate with characteristics of learning experiences in a digital learning environment. Healthy first-year medical students (n = 104) experienced a regular practical course in Microscopic Anatomy either in face-to-face learning, in passive online learning or in an interaction-enhanced version of online learning. Compared to passive online learning, students engaged in the interaction-enhanced version of online learning displayed a significantly reduced Heart Rate Variability (P 0.001, partial η
2 = 0.381) along with a strong increase in salivary cortisol (P 0.001, partial η2 = 0.179) and salivary alpha-amylase activity (P 0.001, partial η2 = 0.195). These results demonstrated that the physiological arousal of students engaged in online learning can be enhanced via interactive teaching methods and pointed towards clear correlations between higher physiological responses and elementary criteria of learning experience such as engagement and attention., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)- Published
- 2023
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27. Induced Endothelial Cell-Integrated Liver Assembloids Promote Hepatic Maturation and Therapeutic Effect on Cholestatic Liver Fibrosis.
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Nam D, Park MR, Lee H, Bae SC, Gerovska D, Araúzo-Bravo MJ, Zaehres H, Schöler HR, and Kim JB
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- Animals, Liver Cirrhosis metabolism, Organoids metabolism, Cholestasis metabolism, Endothelial Cells
- Abstract
The transplantation of pluripotent stem cell (PSC)-derived liver organoids has been studied to solve the current donor shortage. However, the differentiation of unintended cell populations, difficulty in generating multi-lineage organoids, and tumorigenicity of PSC-derived organoids are challenges. However, direct conversion technology has allowed for the generation lineage-restricted induced stem cells from somatic cells bypassing the pluripotent state, thereby eliminating tumorigenic risks. Here, liver assembloids (iHEAs) were generated by integrating induced endothelial cells (iECs) into the liver organoids (iHLOs) generated with induced hepatic stem cells (iHepSCs). Liver assembloids showed enhanced functional maturity compared to iHLOs in vitro and improved therapeutic effects on cholestatic liver fibrosis animals in vivo. Mechanistically, FN1 expressed from iECs led to the upregulation of Itgα5/β1 and Hnf4α in iHEAs and were correlated to the decreased expression of genes related to hepatic stellate cell activation such as Lox and Spp1 in the cholestatic liver fibrosis animals. In conclusion, our study demonstrates the possibility of generating transplantable iHEAs with directly converted cells, and our results evidence that integrating iECs allows iHEAs to have enhanced hepatic maturation compared to iHLOs.
- Published
- 2022
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28. Direct monitoring of live human pluripotent stem cells by a highly selective pluripotency sensor.
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Park MR, Nam D, Lee H, Chang YT, Zaehres H, and Kim JB
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- Cell Differentiation, Humans, Molecular Structure, Fluorescent Dyes chemistry, High-Throughput Screening Assays, Induced Pluripotent Stem Cells cytology
- Abstract
Human pluripotent stem cells (hPSCs) are a useful cell source for regenerative medicine. Despite having a potential of hPSCs for cell-based therapy, there is a need for a selective human pluripotency sensor for monitoring of live hPSCs. Here, we report the discovery of a novel pluripotency sensor (SHI5) from BODIPY-based library by high-throughput cell-based screening and describe the use of SHI5 to identify and isolate human embryonic stem cells and human induced pluripotent stem cells. We demonstrate that SHI5-based assay can be applied to live cells that gain pluripotency in the reprogramming process without any effect on their viability. We also show that SHI5 is internalized through a clathrin-mediated endocytosis pathway. These findings suggest that SHI5 can be an attractive sensor for pluripotency cells during reprogramming. Taken together, SHI5-based screening for hPSCs opens probably unlimited possibilities of detection probe for hPSC therapy via assures their safety issue., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
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29. Sequentially induced motor neurons from human fibroblasts facilitate locomotor recovery in a rodent spinal cord injury model.
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Lee H, Lee HY, Lee BE, Gerovska D, Park SY, Zaehres H, Araúzo-Bravo MJ, Kim JI, Ha Y, Schöler HR, and Kim JB
- Subjects
- Animals, Cell Transplantation, Disease Models, Animal, Female, Humans, LIM-Homeodomain Proteins metabolism, Male, Mice, Mice, Nude, Motor Neurons physiology, Octamer Transcription Factor-3 metabolism, Spinal Cord Injuries physiopathology, Transcription Factors metabolism, Fibroblasts physiology, Gene Expression Regulation, LIM-Homeodomain Proteins genetics, Locomotion physiology, Motor Neurons transplantation, Octamer Transcription Factor-3 genetics, Recovery of Function physiology, Spinal Cord Injuries therapy, Transcription Factors genetics
- Abstract
Generation of autologous human motor neurons holds great promise for cell replacement therapy to treat spinal cord injury (SCI). Direct conversion allows generation of target cells from somatic cells, however, current protocols are not practicable for therapeutic purposes since converted cells are post-mitotic that are not scalable. Therefore, therapeutic effects of directly converted neurons have not been elucidated yet. Here, we show that human fibroblasts can be converted into induced motor neurons (iMNs) by sequentially inducing POU5F1(OCT4) and LHX3 . Our strategy enables scalable production of pure iMNs because of the transient acquisition of proliferative iMN-intermediate cell stage which is distinct from neural progenitors. iMNs exhibited hallmarks of spinal motor neurons including transcriptional profiles, electrophysiological property, synaptic activity, and neuromuscular junction formation. Remarkably, transplantation of iMNs showed therapeutic effects, promoting locomotor functional recovery in rodent SCI model. Together, our advanced strategy will provide tools to acquire sufficient human iMNs that may represent a promising cell source for personalized cell therapy., Competing Interests: HL, HL, BL, DG, SP, HZ, MA, JK, YH, HS, JK No competing interests declared, (© 2020, Lee et al.)
- Published
- 2020
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30. Synapse alterations precede neuronal damage and storage pathology in a human cerebral organoid model of CLN3-juvenile neuronal ceroid lipofuscinosis.
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Gomez-Giro G, Arias-Fuenzalida J, Jarazo J, Zeuschner D, Ali M, Possemis N, Bolognin S, Halder R, Jäger C, Kuper WFE, van Hasselt PM, Zaehres H, Del Sol A, van der Putten H, Schöler HR, and Schwamborn JC
- Subjects
- CRISPR-Cas Systems, Endothelial Cells pathology, Humans, Induced Pluripotent Stem Cells physiology, Lysosomes pathology, Mutation, Organoids, Cerebral Cortex growth & development, Cerebral Cortex pathology, Membrane Glycoproteins genetics, Molecular Chaperones genetics, Neuronal Ceroid-Lipofuscinoses genetics, Neuronal Ceroid-Lipofuscinoses pathology, Neurons pathology, Synapses pathology
- Abstract
The juvenile form of neuronal ceroid Lipofuscinosis (JNCL) is the most common form within this group of rare lysosomal storage disorders, causing pediatric neurodegeneration. The genetic disorder, which is caused by recessive mutations affecting the CLN3 gene, features progressive vision loss, cognitive and motor decline and other psychiatric conditions, seizure episodes, leading to premature death. Animal models have traditionally aid the understanding of the disease mechanisms and pathology and are very relevant for biomarker research and therapeutic testing. Nevertheless, there is a need for establishing reliable and predictive human cellular models to study the disease. Since patient material, particularly from children, is scarce and difficult to obtain, we generated an engineered a CLN3-mutant isogenic human induced pluripotent stem cell (hiPSC) line carrying the c.1054C → T pathologic variant, using state of the art CRISPR/Cas9 technology. To prove the suitability of the isogenic pair to model JNCL, we screened for disease-specific phenotypes in non-neuronal two-dimensional cell culture models as well as in cerebral brain organoids. Our data demonstrates that the sole introduction of the pathogenic variant gives rise to classical hallmarks of JNCL in vitro. Additionally, we discovered an alteration of the splicing caused by this particular mutation. Next, we derived cerebral organoids and used them as a neurodevelopmental model to study the particular effects of the CLN3
Q352X mutation during brain formation in the disease context. About half of the mutation -carrying cerebral organoids completely failed to develop normally. The other half, which escaped this severe defect were used for the analysis of more subtle alterations. In these escapers, whole-transcriptome analysis demonstrated early disease signatures, affecting pathways related to development, corticogenesis and synapses. Complementary metabolomics analysis confirmed decreased levels of cerebral tissue metabolites, some particularly relevant for synapse formation and neurotransmission, such as gamma-amino butyric acid (GABA). Our data suggests that a mutation in CLN3 severely affects brain development. Furthermore, before disease onset, disease -associated neurodevelopmental changes, particular concerning synapse formation and function, occur.- Published
- 2019
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31. Stem cell reprogramming: blood, neurons, and beyond.
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Robin C, Jaffredo T, and Zaehres H
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- Humans, Neurons cytology, Cell Differentiation genetics, Cellular Reprogramming genetics, Neurons metabolism, Stem Cells
- Published
- 2019
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32. Enhanced Ex Vivo Generation of Erythroid Cells from Human Induced Pluripotent Stem Cells in a Simplified Cell Culture System with Low Cytokine Support.
- Author
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Bernecker C, Ackermann M, Lachmann N, Rohrhofer L, Zaehres H, Araúzo-Bravo MJ, van den Akker E, Schlenke P, and Dorn I
- Subjects
- CD36 Antigens genetics, Cell Lineage genetics, Cellular Microenvironment genetics, Cytokines genetics, Erythrocytes cytology, Erythropoiesis genetics, Hematopoietic Stem Cells cytology, Humans, Leukosialin genetics, Cell Culture Techniques methods, Cell Differentiation genetics, Erythroid Cells cytology, Induced Pluripotent Stem Cells cytology
- Abstract
Red blood cell (RBC) differentiation from human induced pluripotent stem cells (hiPSCs) offers great potential for developmental studies and innovative therapies. However, ex vivo erythropoiesis from hiPSCs is currently limited by low efficiency and unphysiological conditions of common culture systems. Especially, the absence of a physiological niche may impair cell growth and lineage-specific differentiation. We here describe a simplified, xeno- and feeder-free culture system for prolonged RBC generation that uses low numbers of supporting cytokines [stem cell factor (SCF), erythropoietin (EPO), and interleukin 3 (IL-3)] and is based on the intermediate development of a "hematopoietic cell forming complex (HCFC)." From this HCFC, CD43
+ hematopoietic cells (purity >95%) were continuously released into the supernatant and could be collected repeatedly over a period of 6 weeks for further erythroid differentiation. The released cells were mainly CD34+ /CD45+ progenitors with high erythroid colony-forming potential and CD36+ erythroid precursors. A total of 1.5 × 107 cells could be harvested from the supernatant of one six-well plate, showing 100- to 1000-fold amplification during subsequent homogeneous differentiation into GPA+ erythroid cells. Mean enucleation rates near 40% (up to 60%) further confirmed the potency of the system. These benefits may be explained by the generation of a niche within the HCFC that mimics the spatiotemporal signaling of the physiological microenvironment in which erythropoiesis occurs. Compared to other protocols, this method provides lower complexity, less cytokine and medium consumption, higher cellular output, and better enucleation. In addition, slight modifications in cytokine addition shift the system toward continuous generation of granulocytes and macrophages.- Published
- 2019
- Full Text
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33. Oct4 and Hnf4α-induced hepatic stem cells ameliorate chronic liver injury in liver fibrosis model.
- Author
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Park MR, Wong MS, Araúzo-Bravo MJ, Lee H, Nam D, Park SY, Seo HD, Lee SM, Zeilhofer HF, Zaehres H, Schöler HR, and Kim JB
- Subjects
- Animals, Male, Mice, Carbon Tetrachloride Poisoning genetics, Carbon Tetrachloride Poisoning metabolism, Carbon Tetrachloride Poisoning therapy, Hepatocyte Nuclear Factor 4 genetics, Hepatocyte Nuclear Factor 4 metabolism, Induced Pluripotent Stem Cells metabolism, Induced Pluripotent Stem Cells pathology, Induced Pluripotent Stem Cells transplantation, Liver metabolism, Liver pathology, Liver Cirrhosis chemically induced, Liver Cirrhosis genetics, Liver Cirrhosis metabolism, Liver Cirrhosis therapy, Lung Injury chemically induced, Lung Injury genetics, Lung Injury metabolism, Octamer Transcription Factor-3 genetics, Octamer Transcription Factor-3 metabolism, Stem Cell Transplantation
- Abstract
Direct conversion from fibroblasts to generate hepatocyte like-cells (iHeps) bypassing the pluripotent state has been described in previous reports as an attractive method acquiring hepatocytes for cell-based therapy. The limited proliferation of iHeps, however, has hampered it uses in cell-based therapy. Since hepatic stem cells (HepSCs) possess self-renewal and bipotency with the capacity to differentiate into both hepatocytes and cholangiocytes, they have therapeutic potential for treating liver disease. Here, we investigated the therapeutic effects of induced HepSCs (iHepSCs) on a carbon tetrachloride (CCl4)-induced liver fibrosis model. We demonstrate that Oct4 and Hnf4a are sufficient to convert fibroblasts into expandable iHepSCs. Hepatocyte-like cells derived from iHepSCs (iHepSC-HEPs) exhibit the typical morphology of hepatocytes and hepatic functions, including glycogen storage, low-density lipoprotein (LDL) uptake, Indocyanine green (ICG) detoxification, drug metabolism, urea production, and albumin secretion. iHepSCs-derived cholangiocyte-like cells (iHepSC-CLCs) expressed cholangiocyte-specific markers and formed cysts and tubule-like structures with apical-basal polarity and secretory function in three-dimensional culture condition. Furthermore, iHepSCs showed anti-inflammatory and anti-fibrotic effects in CCl4-induced liver fibrosis. This study demonstrates that Oct4 and Hnf4α-induced HepSCs show typical hepatic and biliary functionality in vitro. It also presents the therapeutic effect of iHepSCs in liver fibrosis. Therefore, directly converting iHepSCs from somatic cells may facilitate the development of patient-specific cell-based therapy for chronic liver damage., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2019
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34. Novel Tools towards Magnetic Guidance of Neurite Growth: (I) Guidance of Magnetic Nanoparticles into Neurite Extensions of Induced Human Neurons and In Vitro Functionalization with RAS Regulating Proteins.
- Author
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Schöneborn H, Raudzus F, Secret E, Otten N, Michel A, Fresnais J, Ménager C, Siaugue JM, Zaehres H, Dietzel ID, and Heumann R
- Abstract
Parkinson's disease (PD) is a neurodegenerative disease associated with loss or dysfunction of dopaminergic neurons located in the substantia nigra (SN), and there is no cure available. An emerging new approach for treatment is to transplant human induced dopaminergic neurons directly into the denervated striatal brain target region. Unfortunately, neurons grafted into the substantia nigra are unable to grow axons into the striatum and thus do not allow recovery of the original connectivity. Towards overcoming this general limitation in guided neuronal regeneration, we develop here magnetic nanoparticles functionalized with proteins involved in the regulation of axonal growth. We show covalent binding of constitutive active human rat sarcoma (RAS) proteins or RAS guanine nucleotide exchange factor catalytic domain of son of sevenless (SOS) by fluorescence correlation spectroscopy and multiangle light scattering as well as the characterization of exchange factor activity. Human dopaminergic neurons were differentiated from neural precursor cells and characterized by electrophysiological and immune histochemical methods. Furthermore, we demonstrate magnetic translocation of cytoplasmic γ-Fe
2 O3 @SiO2 core-shell nanoparticles into the neurite extensions of induced human neurons. Altogether, we developed tools towards remote control of directed neurite growth in human dopaminergic neurons. These results may have relevance for future therapeutic approaches of cell replacement therapy in Parkinson's disease.- Published
- 2019
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35. bHLH Transcription Factor Math6 Antagonizes TGF-β Signalling in Reprogramming, Pluripotency and Early Cell Fate Decisions.
- Author
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Divvela SSK, Nell P, Napirei M, Zaehres H, Chen J, Gerding WM, Nguyen HP, Gao S, and Brand-Saberi B
- Subjects
- Animals, Basic Helix-Loop-Helix Transcription Factors genetics, Epithelium metabolism, Female, Gene Expression Regulation, Developmental, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Male, Mesoderm metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Mouse Embryonic Stem Cells metabolism, Pluripotent Stem Cells metabolism, Basic Helix-Loop-Helix Transcription Factors metabolism, Cell Lineage, Cellular Reprogramming, Pluripotent Stem Cells cytology, Signal Transduction, Transforming Growth Factor beta metabolism
- Abstract
The basic helix-loop-helix (bHLH) transcription factor Math6 (Atonal homolog 8; Atoh8) plays a crucial role in a number of cellular processes during embryonic development, iron metabolism and tumorigenesis. We report here on its involvement in cellular reprogramming from fibroblasts to induced pluripotent stem cells, in the maintenance of pluripotency and in early fate decisions during murine development. Loss of Math6 disrupts mesenchymal-to-epithelial transition during reprogramming and primes pluripotent stem cells towards the mesendodermal fate. Math6 can thus be considered a regulator of reprogramming and pluripotent stem cell fate. Additionally, our results demonstrate the involvement of Math6 in SMAD-dependent TGF beta signalling. We furthermore monitor the presence of the Math6 protein during these developmental processes using a newly generated Math6Flag-tag mouse. Taken together, our results suggest that Math6 counteracts TGF beta signalling and, by this, affects the initiating step of cellular reprogramming, as well as the maintenance of pluripotency and early differentiation., Competing Interests: The authors declare no competing interests.
- Published
- 2019
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36. The in vivo timeline of differentiation of engrafted human neural progenitor cells.
- Author
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Vogel S, Schäfer C, Hess S, Folz-Donahue K, Nelles M, Minassian A, Schwarz MK, Kukat C, Ehrlich M, Zaehres H, Kloppenburg P, Hoehn M, and Aswendt M
- Subjects
- Animals, Cells, Cultured, Humans, Male, Mice, Neurogenesis, Astrocytes cytology, Cell Differentiation, Cell Lineage, Induced Pluripotent Stem Cells cytology, Neural Stem Cells cytology, Neurons cytology, Oligodendroglia cytology
- Abstract
Understanding the individual timeline of stem cell differentiation in vivo is critical for evaluating stem cell properties in animal models. However, with conventional ex vivo techniques, such as histology, the individual timeline of differentiation is not accessible. Therefore, we designed lentiviral plasmids with cell-specific promoters to control the expression of bioluminescence and fluorescence imaging reporters. Promoter-dependent reporter expression in transduced human induced pluripotent stem cell-derived neural progenitor cells (hNPCs) was an effective indicator of differentiation in cell culture. A 12-week in vivo imaging observation period revealed the time profile of differentiation of engrafted hNPCs in the mouse brain into astrocytes and mature neurons which was verified by immunostainings, patch-clamp electrophysiology, and light-sheet fluorescence microscopy. The lentiviral vectors validated in this study provide an efficient imaging toolbox for non-invasive and longitudinal characterization of stem cell differentiation, in vitro screenings, and in vivo studies of cell therapy in animal models., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2019
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37. The Hinrichsen Embryology Collection: Digitization of Historical Histological Human Embryonic Slides and MRI of Whole Fetuses.
- Author
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Maricic N, Khaveh N, Marheinecke C, Wald J, Helluy X, Liermann D, Zaehres H, and Brand-Saberi B
- Subjects
- Embryo, Mammalian abnormalities, Female, Fetus abnormalities, Humans, Infant, Newborn, Male, Embryo, Mammalian pathology, Fetus diagnostic imaging, Imaging, Three-Dimensional, Magnetic Resonance Imaging
- Abstract
The number of human embryology collections is very limited worldwide. Some of these comprise the Carnegie Collection, Kyoto Collection, and the Blechschmidt Collection. One further embryonic collection is the Hinrichsen Collection of the Ruhr University Bochum, Germany, which also contains very well-preserved embryos/fetuses, along with approximately 16,000 histological sections. The digitization of this collection is indispensable to enable conservation of the collection for the future and to provide a large group of embryologists, researchers, and physicians access to these histological slides. A small selection of these scans is available at the website of the Digital Embryology Consortium [https://-human-embryology.org/wiki/Main_Page]., (© 2019 S. Karger AG, Basel.)
- Published
- 2019
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38. FACS-Assisted CRISPR-Cas9 Genome Editing Facilitates Parkinson's Disease Modeling.
- Author
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Arias-Fuenzalida J, Jarazo J, Qing X, Walter J, Gomez-Giro G, Nickels SL, Zaehres H, Schöler HR, and Schwamborn JC
- Subjects
- Alleles, Cells, Cultured, DNA End-Joining Repair genetics, Flow Cytometry methods, Humans, Induced Pluripotent Stem Cells cytology, CRISPR-Cas Systems, Gene Editing methods, Induced Pluripotent Stem Cells metabolism, Parkinson Disease genetics, alpha-Synuclein genetics
- Abstract
Genome editing and human induced pluripotent stem cells hold great promise for the development of isogenic disease models and the correction of disease-associated mutations for isogenic tissue therapy. CRISPR-Cas9 has emerged as a versatile and simple tool for engineering human cells for such purposes. However, the current protocols to derive genome-edited lines require the screening of a great number of clones to obtain one free of random integration or on-locus non-homologous end joining (NHEJ)-containing alleles. Here, we describe an efficient method to derive biallelic genome-edited populations by the use of fluorescent markers. We call this technique FACS-assisted CRISPR-Cas9 editing (FACE). FACE allows the derivation of correctly edited polyclones carrying a positive selection fluorescent module and the exclusion of non-edited, random integrations and on-target allele NHEJ-containing cells. We derived a set of isogenic lines containing Parkinson's-disease-associated mutations in α-synuclein and present their comparative phenotypes., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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39. GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells.
- Author
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van der Wal E, Bergsma AJ, van Gestel TJM, In 't Groen SLM, Zaehres H, Araúzo-Bravo MJ, Schöler HR, van der Ploeg AT, and Pijnappel WWMP
- Abstract
Pompe disease is a metabolic myopathy caused by deficiency of the acid α-glucosidase (GAA) enzyme and results in progressive wasting of skeletal muscle cells. The c.-32-13T>G (IVS1) GAA variant promotes exon 2 skipping during pre-mRNA splicing and is the most common variant for the childhood/adult disease form. We previously identified antisense oligonucleotides (AONs) that promoted GAA exon 2 inclusion in patient-derived fibroblasts. It was unknown how these AONs would affect GAA splicing in skeletal muscle cells. To test this, we expanded induced pluripotent stem cell (iPSC)-derived myogenic progenitors and differentiated these to multinucleated myotubes. AONs restored splicing in myotubes to a similar extent as in fibroblasts, suggesting that they act by modulating the action of shared splicing regulators. AONs targeted the putative polypyrimidine tract of a cryptic splice acceptor site that was part of a pseudo exon in GAA intron 1. Blocking of the cryptic splice donor of the pseudo exon with AONs likewise promoted GAA exon 2 inclusion. The simultaneous blocking of the cryptic acceptor and cryptic donor sites restored the majority of canonical splicing and alleviated GAA enzyme deficiency. These results highlight the relevance of cryptic splicing in human disease and its potential as therapeutic target for splicing modulation using AONs., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
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40. Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells.
- Author
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Kessel KU, Bluemke A, Schöler HR, Zaehres H, Schlenke P, and Dorn I
- Abstract
Background: The ex vivo generation of human hematopoietic stem cells (HSCs) with long-term repopulating capacity and multi-lineage differentiation potential represents the holy grail of hematopoiesis research. In principle, human induced pluripotent stem cells (hiPSCs) provide the tool for both studying molecular mechanisms of hematopoietic development and the ex vivo production of 'true' HSCs for transplantation purposes and lineage-specific cells, e.g. red blood cells, for transfusion purposes. CD43-expressing cells have been reported as the first hematopoietic cells during development, but whether or not these possess multilineage differentiation and long-term engraftment potential is incompletely understood., Methods: We performed ex vivo generation of hematopoietic cells from hiPSCs using an embryoid body(EB)-based, xeno-product-free differentiation protocol. We investigated the multilineage differentiation potential of different FACS-sorted CD43-expressing cell subsets by colony-forming assays in semisolid media. Further, erythroid differentiation was investigated in more detail using established protocols., Results: By using CD43, we were able to measure hematopoietic induction efficiency during hiPSC-derived EB differentiation. Further, we determined CD43+ cells as the cell population of origin for in vitro erythropoiesis. Furthermore, colony formation demonstrates that the multipotent hematopoietic stem and progenitor cell fraction is particulary enriched in the CD43
hi CD45+ population.- Published
- 2017
- Full Text
- View/download PDF
41. Rapid and efficient generation of oligodendrocytes from human induced pluripotent stem cells using transcription factors.
- Author
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Ehrlich M, Mozafari S, Glatza M, Starost L, Velychko S, Hallmann AL, Cui QL, Schambach A, Kim KP, Bachelin C, Marteyn A, Hargus G, Johnson RM, Antel J, Sterneckert J, Zaehres H, Schöler HR, Baron-Van Evercooren A, and Kuhlmann T
- Subjects
- Animals, Biomarkers, Brain metabolism, Brain pathology, Brain ultrastructure, Cell Death genetics, Cell Lineage genetics, Cells, Cultured, Cluster Analysis, Demyelinating Diseases genetics, Demyelinating Diseases metabolism, Demyelinating Diseases pathology, Disease Models, Animal, Ectopic Gene Expression, Gene Expression Profiling, Humans, Mice, Mutation, Myelin Basic Protein genetics, Myelin Basic Protein metabolism, Myelin Sheath genetics, Myelin Sheath metabolism, Neural Stem Cells cytology, Neural Stem Cells metabolism, Oxidative Stress, Spinal Cord metabolism, Spinal Cord pathology, Spinal Cord ultrastructure, Transcription Factors metabolism, Transcriptome, tau Proteins genetics, tau Proteins metabolism, Cell Differentiation genetics, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Oligodendroglia cytology, Oligodendroglia metabolism, Transcription Factors genetics
- Abstract
Rapid and efficient protocols to generate oligodendrocytes (OL) from human induced pluripotent stem cells (iPSC) are currently lacking, but may be a key technology to understand the biology of myelin diseases and to develop treatments for such disorders. Here, we demonstrate that the induction of three transcription factors (SOX10, OLIG2, NKX6.2) in iPSC-derived neural progenitor cells is sufficient to rapidly generate O4
+ OL with an efficiency of up to 70% in 28 d and a global gene-expression profile comparable to primary human OL. We further demonstrate that iPSC-derived OL disperse and myelinate the CNS of Mbpshi/shi Rag-/- mice during development and after demyelination, are suitable for in vitro myelination assays, disease modeling, and screening of pharmacological compounds potentially promoting oligodendroglial differentiation. Thus, the strategy presented here to generate OL from iPSC may facilitate the studying of human myelin diseases and the development of high-throughput screening platforms for drug discovery.- Published
- 2017
- Full Text
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42. Astrocyte pathology in a human neural stem cell model of frontotemporal dementia caused by mutant TAU protein.
- Author
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Hallmann AL, Araúzo-Bravo MJ, Mavrommatis L, Ehrlich M, Röpke A, Brockhaus J, Missler M, Sterneckert J, Schöler HR, Kuhlmann T, Zaehres H, and Hargus G
- Subjects
- Annexin A2 metabolism, Astrocytes cytology, Astrocytes pathology, Cell Differentiation, Coculture Techniques, Frontotemporal Dementia genetics, Humans, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Models, Biological, Neural Stem Cells cytology, Neural Stem Cells metabolism, Neurons cytology, Neurons metabolism, Oxidative Stress, Polymorphism, Single Nucleotide, Protein Isoforms genetics, Transcriptome, Ubiquitination, Astrocytes metabolism, Frontotemporal Dementia pathology, tau Proteins genetics
- Abstract
Astroglial pathology is seen in various neurodegenerative diseases including frontotemporal dementia (FTD), which can be caused by mutations in the gene encoding the microtubule-associated protein TAU (MAPT). Here, we applied a stem cell model of FTD to examine if FTD astrocytes carry an intrinsic propensity to degeneration and to determine if they can induce non-cell-autonomous effects in neighboring neurons. We utilized CRISPR/Cas9 genome editing in human induced pluripotent stem (iPS) cell-derived neural progenitor cells (NPCs) to repair the FTD-associated N279K MAPT mutation. While astrocytic differentiation was not impaired in FTD NPCs derived from one patient carrying the N279K MAPT mutation, FTD astrocytes appeared larger, expressed increased levels of 4R-TAU isoforms, demonstrated increased vulnerability to oxidative stress and elevated protein ubiquitination and exhibited disease-associated changes in transcriptome profiles when compared to astrocytes derived from one control individual and to the isogenic control. Interestingly, co-culture experiments with FTD astrocytes revealed increased oxidative stress and robust changes in whole genome expression in previously healthy neurons. Our study highlights the utility of iPS cell-derived NPCs to elucidate the role of astrocytes in the pathogenesis of FTD.
- Published
- 2017
- Full Text
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43. The development of anatomy: from macroscopic body dissections to stem cell-derived organoids.
- Author
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Brand-Saberi B and Zaehres H
- Subjects
- Anatomy trends, Animals, Cell Differentiation, Humans, Organoids metabolism, Pluripotent Stem Cells metabolism, Anatomy methods, Organoids cytology, Pluripotent Stem Cells cytology
- Abstract
Anatomy as a descriptive topic of research and instruction in medicine has been increasingly influenced by discoveries in molecular cell and developmental biology and most recently the advent of human induced pluripotent stem cells and organoids. We summarize here how anatomy has been influenced by developmental and stem cell biologists, and how in vitro modelling of the three-dimensional body environment is emerging to understand structure and function of cells during differentiation processes in development and disease.
- Published
- 2016
- Full Text
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44. Comparative transcriptome analysis in induced neural stem cells reveals defined neural cell identities in vitro and after transplantation into the adult rodent brain.
- Author
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Hallmann AL, Araúzo-Bravo MJ, Zerfass C, Senner V, Ehrlich M, Psathaki OE, Han DW, Tapia N, Zaehres H, Schöler HR, Kuhlmann T, and Hargus G
- Subjects
- Animals, Brain pathology, Cell Differentiation, Cells, Cultured, Cellular Reprogramming, Cluster Analysis, Fibroblasts cytology, Gene Expression Profiling, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells transplantation, Mice, Mice, Inbred C57BL, Neural Stem Cells cytology, Neural Stem Cells transplantation, Transcription Factors genetics, Transcription Factors metabolism, Brain metabolism, Induced Pluripotent Stem Cells metabolism, Neural Stem Cells metabolism, Transcriptome
- Abstract
Reprogramming technology enables the production of neural progenitor cells (NPCs) from somatic cells by direct transdifferentiation. However, little is known on how neural programs in these induced neural stem cells (iNSCs) differ from those of alternative stem cell populations in vitro and in vivo. Here, we performed transcriptome analyses on murine iNSCs in comparison to brain-derived neural stem cells (NSCs) and pluripotent stem cell-derived NPCs, which revealed distinct global, neural, metabolic and cell cycle-associated marks in these populations. iNSCs carried a hindbrain/posterior cell identity, which could be shifted towards caudal, partially to rostral but not towards ventral fates in vitro. iNSCs survived after transplantation into the rodent brain and exhibited in vivo-characteristics, neural and metabolic programs similar to transplanted NSCs. However, iNSCs vastly retained caudal identities demonstrating cell-autonomy of regional programs in vivo. These data could have significant implications for a variety of in vitro- and in vivo-applications using iNSCs., (Copyright © 2016 Roslin Cells Ltd. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
45. Establishment of feeder-free culture system for human induced pluripotent stem cell on DAS nanocrystalline graphene.
- Author
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Lee H, Nam D, Choi JK, Araúzo-Bravo MJ, Kwon SY, Zaehres H, Lee T, Park CY, Kang HW, Schöler HR, and Kim JB
- Subjects
- Cell Adhesion, Cell Differentiation, Cell Line, Cell Proliferation, Feeder Cells cytology, Gene Expression, Humans, Induced Pluripotent Stem Cells metabolism, Surface Properties, Cell Culture Techniques methods, Graphite chemistry, Induced Pluripotent Stem Cells cytology, Nanostructures chemistry
- Abstract
The maintenance of undifferentiated human pluripotent stem cells (hPSC) under xeno-free condition requires the use of human feeder cells or extracellular matrix (ECM) coating. However, human-derived sources may cause human pathogen contamination by viral or non-viral agents to the patients. Here we demonstrate feeder-free and xeno-free culture system for hPSC expansion using diffusion assisted synthesis-grown nanocrystalline graphene (DAS-NG), a synthetic non-biological nanomaterial which completely rule out the concern of human pathogen contamination. DAS-NG exhibited advanced biocompatibilities including surface nanoroughness, oxygen containing functional groups and hydrophilicity. hPSC cultured on DAS-NG could maintain pluripotency in vitro and in vivo, and especially cell adhesion-related gene expression profile was comparable to those of cultured on feeders, while hPSC cultured without DAS-NG differentiated spontaneously with high expression of somatic cell-enriched adhesion genes. This feeder-free and xeno-free culture method using DAS-NG will facilitate the generation of clinical-grade hPSC.
- Published
- 2016
- Full Text
- View/download PDF
46. Factor-Reduced Human Induced Pluripotent Stem Cells Efficiently Differentiate into Neurons Independent of the Number of Reprogramming Factors.
- Author
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Hermann A, Kim JB, Srimasorn S, Zaehres H, Reinhardt P, Schöler HR, and Storch A
- Abstract
Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) by overexpression of the transcription factors OCT4, SOX2, KLF4, and c-Myc holds great promise for the development of personalized cell replacement therapies. In an attempt to minimize the risk of chromosomal disruption and to simplify reprogramming, several studies demonstrated that a reduced set of reprogramming factors is sufficient to generate iPSC. We recently showed that a reduction of reprogramming factors in murine cells not only reduces reprogramming efficiency but also may worsen subsequent differentiation. To prove whether this is also true for human cells, we compared the efficiency of neuronal differentiation of iPSC generated from fetal human neural stem cells with either one (OCT4; hiPSC1F-NSC) or two (OCT4, KLF4; hiPSC2F-NSC) reprogramming factors with iPSC produced from human fibroblasts using three (hiPSC3F-FIB) or four reprogramming factors (hiPSC4F-FIB). After four weeks of coculture with PA6 stromal cells, neuronal differentiation of hiPSC1F-NSC and hiPSC2F-NSC was as efficient as iPSC3F-FIB or iPSC4F-FIB. We conclude that a reduction of reprogramming factors in human cells does reduce reprogramming efficiency but does not alter subsequent differentiation into neural lineages. This is of importance for the development of future application of iPSC in cell replacement therapies.
- Published
- 2016
- Full Text
- View/download PDF
47. Oct4-induced oligodendrocyte progenitor cells enhance functional recovery in spinal cord injury model.
- Author
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Kim JB, Lee H, Araúzo-Bravo MJ, Hwang K, Nam D, Park MR, Zaehres H, Park KI, and Lee SJ
- Subjects
- Animals, Cells, Cultured, Disease Models, Animal, Fibroblasts cytology, Immunohistochemistry, Karyotype, Male, Mice, Mice, SCID, Octamer Transcription Factor-3 genetics, Oligodendroglia cytology, Rats, Recovery of Function physiology, Spinal Cord Injuries genetics, Stem Cell Transplantation, Stem Cells cytology, Stem Cells physiology, Octamer Transcription Factor-3 metabolism, Oligodendroglia metabolism, Oligodendroglia physiology, Spinal Cord Injuries metabolism, Spinal Cord Injuries therapy, Stem Cells metabolism
- Abstract
The generation of patient-specific oligodendrocyte progenitor cells (OPCs) holds great potential as an expandable cell source for cell replacement therapy as well as drug screening in spinal cord injury or demyelinating diseases. Here, we demonstrate that induced OPCs (iOPCs) can be directly derived from adult mouse fibroblasts by Oct4-mediated direct reprogramming, using anchorage-independent growth to ensure high purity. Homogeneous iOPCs exhibit typical small-bipolar morphology, maintain their self-renewal capacity and OPC marker expression for more than 31 passages, share high similarity in the global gene expression profile to wild-type OPCs, and give rise to mature oligodendrocytes and astrocytes in vitro and in vivo. Notably, transplanted iOPCs contribute to functional recovery in a spinal cord injury (SCI) model without tumor formation. This study provides a simple strategy to generate functional self-renewing iOPCs and yields insights for the in-depth study of demyelination and regenerative medicine., (© 2015 The Authors.)
- Published
- 2015
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48. Distinct Neurodegenerative Changes in an Induced Pluripotent Stem Cell Model of Frontotemporal Dementia Linked to Mutant TAU Protein.
- Author
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Ehrlich M, Hallmann AL, Reinhardt P, Araúzo-Bravo MJ, Korr S, Röpke A, Psathaki OE, Ehling P, Meuth SG, Oblak AL, Murrell JR, Ghetti B, Zaehres H, Schöler HR, Sterneckert J, Kuhlmann T, and Hargus G
- Subjects
- Frontotemporal Dementia pathology, Gene Expression Profiling, Gene Expression Regulation, Humans, Induced Pluripotent Stem Cells metabolism, Mitochondria metabolism, Mitochondria pathology, Mutation, Neurites pathology, Oxidative Stress genetics, Unfolded Protein Response genetics, tau Proteins biosynthesis, Cell Differentiation genetics, Frontotemporal Dementia genetics, Induced Pluripotent Stem Cells pathology, tau Proteins genetics
- Abstract
Frontotemporal dementia (FTD) is a frequent form of early-onset dementia and can be caused by mutations in MAPT encoding the microtubule-associated protein TAU. Because of limited availability of neural cells from patients' brains, the underlying mechanisms of neurodegeneration in FTD are poorly understood. Here, we derived induced pluripotent stem cells (iPSCs) from individuals with FTD-associated MAPT mutations and differentiated them into mature neurons. Patient iPSC-derived neurons demonstrated pronounced TAU pathology with increased fragmentation and phospho-TAU immunoreactivity, decreased neurite extension, and increased but reversible oxidative stress response to inhibition of mitochondrial respiration. Furthermore, FTD neurons showed an activation of the unfolded protein response, and a transcriptome analysis demonstrated distinct, disease-associated gene expression profiles. These findings indicate distinct neurodegenerative changes in FTD caused by mutant TAU and highlight the unique opportunity to use neurons differentiated from patient-specific iPSCs to identify potential targets for drug screening purposes and therapeutic intervention., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
49. DNA damage response in neonatal and adult stromal cells compared with induced pluripotent stem cells.
- Author
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Liedtke S, Biebernick S, Radke TF, Stapelkamp D, Coenen C, Zaehres H, Fritz G, and Kogler G
- Subjects
- Adult, Ataxia Telangiectasia Mutated Proteins metabolism, Drug Evaluation, Preclinical methods, Female, Histones metabolism, Humans, Induced Pluripotent Stem Cells cytology, Infant, Newborn, Male, Phosphorylation drug effects, Stromal Cells cytology, Stromal Cells metabolism, X-Rays, Alkylating Agents pharmacology, Cell Differentiation drug effects, DNA Damage, Induced Pluripotent Stem Cells metabolism, Methylnitrosourea pharmacology, Osteogenesis drug effects
- Abstract
Unlabelled: Comprehensive analyses comparing individual DNA damage response (DDR) of induced pluripotent stem cells (iPSCs) with neonatal stromal cells with respect to their developmental age are limited. The imperative necessity of providing developmental age-matched cell sources for meaningful toxicological drug safety assessments in replacement of animal-based testing strategies is evident. Here, DDR after radiation or treatment with N-methyl-N-nitrosurea (MNU) was determined in iPSCs compared with neonatal and bone marrow stromal cells. Neonatal and adult stromal cells showed no significant morphologically detectable cytotoxicity following treatment with 1 Gy or 1 mM MNU, whereas iPSCs revealed a much higher sensitivity. Foci analyses revealed an effective DNA repair in stromal cell types and iPSCs, as reflected by a rapid formation and disappearance of phosphorylated ATM and γH2AX foci. Furthermore, quantitative polymerase chain reaction analyses revealed the highest basic expression level of DDR and repair-associated genes in iPSCs, followed by neonatal stromal cells and adult stromal cells with the lowest expression levels. In addition, the influence of genotoxic stress prior to and during osteogenic differentiation of neonatal and adult stromal cells was analyzed applying common differentiation procedures. Experiments presented here suggest a developmental age-dependent basic expression level of genes involved in the processing of DNA damage. In addition a differentiation-dependent downregulation of repair genes was observed during osteogenesis. These results strongly support the requirement to provide adequate cell sources for toxicological in vitro drug testing strategies that match to the developmental age and differentiation status of the presumptive target cell of interest., Significance: The results obtained in this study advance the understanding of DNA damage processing in human neonatal stromal cells as compared with adult stromal cells and induced pluripotent stem cells (iPSCs). The data suggest developmental age-dependent differences in DNA damage repair capacity. In iPSCs (closest to embryonic stem cells), the highest expression level of DNA damage response and repair genes was found, followed by neonatal stromal cells and adult stromal cells with the lowest overall expression. In addition, a differentiation-dependent downregulation of repair capacity was observed during osteogenic differentiation in neonatal stromal cells. Notably, the impact of genotoxic stress on osteogenic differentiation depended on the time the genotoxic insult took place and, moreover, was agent-specific. These results strongly support the necessity of offering and establishing adequate cell sources for informative toxicological testing matching to the developmental age and differentiation status of the respective cell of interest., (©AlphaMed Press.)
- Published
- 2015
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50. ERRATUM: Establishment of a primed pluripotent epiblast stem cell in FGF4-based conditions.
- Author
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Joo JY, Choi HW, Kim MJ, Zaehres H, Tapia N, Stehling M, Jung KS, Tae Do J, and Schöler HR
- Published
- 2015
- Full Text
- View/download PDF
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