11 results on '"Hong, Hyenjong"'
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2. A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells
- Author
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Chen, Yu, Tristan, Carlos A., Chen, Lu, Jovanovic, Vukasin M., Malley, Claire, Chu, Pei-Hsuan, Ryu, Seungmi, Deng, Tao, Ormanoglu, Pinar, Tao, Dingyin, Fang, Yuhong, Slamecka, Jaroslav, Hong, Hyenjong, LeClair, Christopher A., Michael, Sam, Austin, Christopher P., Simeonov, Anton, and Singeç, Ilyas more...
- Published
- 2021
- Full Text
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Catalog
3. Suppression of induced pluripotent stem cell generation by the p53-p21 pathway
- Author
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Hong, Hyenjong, Takahashi, Kazutoshi, Ichisaka, Tomoko, Aoi, Takashi, Kanagawa, Osami, Nakagawa, Masato, Okita, Keisuke, and Yamanaka, Shinya
- Subjects
Gene mutations -- Research -- Physiological aspects -- Genetic aspects ,Stem cells -- Research -- Physiological aspects -- Genetic aspects ,Fibroblasts -- Research -- Genetic aspects -- Physiological aspects ,Cellular signal transduction -- Research -- Physiological aspects -- Genetic aspects ,Environmental issues ,Science and technology ,Zoology and wildlife conservation ,Physiological aspects ,Research ,Genetic aspects - Abstract
Induced pluripotent stem(iPS) cells can be generated from somatic cells by the introduction of Oct3/4 (also known as Pou5f1), Sox2, Klf4 and c-Myc, in mouse (1-4) and in human (5-8). The efficiency of this process, however, is low (9). Pluripotency can be induced without c-Myc, but with even lower efficiency (10, 11). A p53 (also known as TP53 in humans and Trp53 in mice) short-interfering RNA (siRNA) was recently shown to promote human iPS cell generation (12), but the specificity and mechanisms remain to be determined. Here we report that up to 10% of transduced mouse embryonic fibroblasts lacking p53 became iPS cells, even without the Myc retrovirus. The p53 deletion also promoted the induction of integration-free mouse iPS cells with plasmid transfection. Furthermore, in the p53-null background, iPS cells were generated from terminally differentiated T lymphocytes. The suppression of p53 also increased the efficiency of humani PS cell generation. DNA microarray analyses identified 34 p53-regulated genes that are common in mouse and human fibroblasts. Functional analyses of these genes demonstrate that the p53-p21 pathway serves as a barrier not only in tumorigenicity, but also in iPS cell generation., We used the Nanog-GFP (green fluorescent protein) reporter system for sensitive and specific identification of iPS cells (3). When the three factors (Oct3/4, Sox2 and Klf4) devoid of c-Myc were [...] more...
- Published
- 2009
4. p53-p21経路によるiPS細胞の樹立抑制
- Author
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Hong, Hyenjong, 篠原, 隆司, 中畑, 龍俊, and 中辻, 憲夫
- Published
- 2012
5. Standards for Deriving Nonhuman Primate-Induced Pluripotent Stem Cells, Neural Stem Cells and Dopaminergic Lineage.
- Author
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Yang, Guang, Hong, Hyenjong, Torres, April, Malloy, Kristen E., Choudhury, Gourav R., Kim, Jeffrey, and Daadi, Marcel M.
- Subjects
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PLURIPOTENT stem cells , *NEURAL stem cells , *DOPAMINERGIC neurons , *CELL culture , *COMMUNICABLE disease immunology - Abstract
Humans and nonhuman primates (NHP) are similar in behavior and in physiology, specifically the structure, function, and complexity of the immune system. Thus, NHP models are desirable for pathophysiology and pharmacology/toxicology studies. Furthermore, NHP-derived induced pluripotent stem cells (iPSCs) may enable transformative developmental, translational, or evolutionary studies in a field of inquiry currently hampered by the limited availability of research specimens. NHP-iPSCs may address specific questions that can be studied back and forth between in vitro cellular assays and in vivo experimentations, an investigational process that in most cases cannot be performed on humans because of safety and ethical issues. The use of NHP model systems and cell specific in vitro models is evolving with iPSC-based three-dimensional (3D) cell culture systems and organoids, which may offer reliable in vitro models and reduce the number of animals used in experimental research. IPSCs have the potential to give rise to defined cell types of any organ of the body. However, standards for deriving defined and validated NHP iPSCs are missing. Standards for deriving high-quality iPSC cell lines promote rigorous and replicable scientific research and likewise, validated cell lines reduce variability and discrepancies in results between laboratories. We have derived and validated NHP iPSC lines by confirming their pluripotency and propensity to differentiate into all three germ layers (ectoderm, mesoderm, and endoderm) according to standards and measurable limits for a set of marker genes. The iPSC lines were characterized for their potential to generate neural stem cells and to differentiate into dopaminergic neurons. These iPSC lines are available to the scientific community. NHP-iPSCs fulfill a unique niche in comparative genomics to understand gene regulatory principles underlying emergence of human traits, in infectious disease pathogenesis, in vaccine development, and in immunological barriers in regenerative medicine. [ABSTRACT FROM AUTHOR] more...
- Published
- 2018
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6. Stress-free cell aggregation by using the CEPT cocktail enhances embryoid body and organoid fitness.
- Author
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Ryu S, Weber C, Chu PH, Ernest B, Jovanovic VM, Deng T, Slamecka J, Hong H, Jethmalani Y, Baskir HM, Inman J, Braisted J, Hirst MB, Simeonov A, Voss TC, Tristan CA, and Singeç I
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- Humans, Reproducibility of Results, Organoids, Cell Differentiation, Embryoid Bodies metabolism, Pluripotent Stem Cells
- Abstract
Embryoid bodies (EBs) and self-organizing organoids derived from human pluripotent stem cells (hPSCs) recapitulate tissue development in a dish and hold great promise for disease modeling and drug development. However, current protocols are hampered by cellular stress and apoptosis during cell aggregation, resulting in variability and impaired cell differentiation. Here, we demonstrate that EBs and various organoid models (e.g., brain, gut, kidney) can be optimized by using the small molecule cocktail named CEPT (chroman 1, emricasan, polyamines, trans-ISRIB), a polypharmacological approach that ensures cytoprotection and cell survival. Application of CEPT for just 24 h during cell aggregation has long-lasting consequences affecting morphogenesis, gene expression, cellular differentiation, and organoid function. Various qualification methods confirmed that CEPT treatment enhanced experimental reproducibility and consistently improved EB and organoid fitness as compared to the widely used ROCK inhibitor Y-27632. Collectively, we discovered that stress-free cell aggregation and superior cell survival in the presence of CEPT are critical quality control determinants that establish a robust foundation for bioengineering complex tissue and organ models., (Creative Commons Attribution license.) more...
- Published
- 2023
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7. Isolation and Differentiation of Self-Renewable Neural Stem Cells from Marmoset-Induced Pluripotent Stem Cells.
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Hong H, Roy-Choudhury G, Kim J, and Daadi MM
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- Animals, Callithrix, Cell Culture Techniques, Cell Differentiation, Cell Self Renewal, Cell Separation methods, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Neural Stem Cells cytology, Neural Stem Cells metabolism
- Abstract
Neural stem cells (NSCs) are multipotent and self-renewing precursor cells that give rise to all cell types of the central nervous system (CNS). They can be used for modeling CNS in vitro, for developmental studies and for cell replacement therapies. NSCs can be derived from pluripotent stem cells through differentiation using specific growth factors. Nonhuman primates (NHP) are critical preclinical models for translational research. Induced pluripotent stem cells (iPSCs) can be generated from NHP for the purposes of allogenic or autologous cell replacement studies. Here, we describe the derivation of NSCs from NHP iPSCs. more...
- Published
- 2019
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8. Reference Transcriptome for Deriving Marmoset Induced Pluripotent Stem Cells.
- Author
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Yang G, Hong H, Torres A, Malloy KE, Roy-Choudhury G, Kim J, and Daadi MM
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- Animals, Biopsy, Callithrix, Cell Line, Embryoid Bodies metabolism, Skin cytology, Skin metabolism, Workflow, Gene Expression Profiling methods, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Regenerative Medicine, Transcriptome
- Abstract
Limited access to primary tissue from various nonhuman primate (NHP) species represents a significant unmet need that hampers progress in understanding unique cellular diversity and gene regulation of specific tissues and organs in stem cell translational research. Most comparative biology studies have been limited to using postmortem tissue usually frozen specimens with limited utility for research. The generation of induced pluripotent stem cell (iPSC) lines from somatic cells, such as adult skin or blood cells, offers an alternative to invasive and ethically controversial interventions for acquiring tissue. Pluripotent iPSCs have virtually an unlimited capacity to proliferate and differentiate into all cell types of the body. We are generating high-quality validated NHP iPSC lines to offer to scientific community and facilitate their research programs. We use the non-integrative episomal vector system to generate iPSCs from NHP skin biopsies. In this chapter we describe the validation of NHP iPSC lines by confirming pluripotency and their propensity to differentiate into all three germ layers ectoderm, mesoderm, and endoderm according to established standards and measurable limits for a set of marker genes incorporated into a scorecard. more...
- Published
- 2019
- Full Text
- View/download PDF
9. Generating Neural Stem Cells from iPSCs with Dopaminergic Neurons Reporter Gene.
- Author
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Hong H and Daadi MM
- Subjects
- CRISPR-Cas Systems, Cell Culture Techniques, Gene Editing, HEK293 Cells, Humans, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Parkinson Disease metabolism, RNA, Guide, CRISPR-Cas Systems, Tyrosine 3-Monooxygenase genetics, Tyrosine 3-Monooxygenase metabolism, Cell Differentiation, Dopaminergic Neurons cytology, Dopaminergic Neurons metabolism, Gene Expression, Genes, Reporter, Neural Stem Cells cytology, Neural Stem Cells metabolism
- Abstract
Genetic reporters offer attractive approaches to investigate gene expression, gene function, and spatiotemporal assessment in vitro and in vivo. Tyrosine hydroxylase (TH) is the rate-limiting enzyme for the biosynthesis of the dopamine neurotransmitter, and thus it has been used as a reliable marker for dopaminergic neurons in vitro and in vivo. Herein we describe a method for making iPSC lines with TH-green fluorescent protein reporter gene using CRISPR/Cas9 technique. more...
- Published
- 2019
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10. A more efficient method to generate integration-free human iPS cells.
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Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S, Hong H, Nakagawa M, Tanabe K, Tezuka K, Shibata T, Kunisada T, Takahashi M, Takahashi J, Saji H, and Yamanaka S
- Subjects
- Asian People genetics, Electroporation, Gene Expression Profiling, Gene Frequency, Genetic Vectors, HLA Antigens genetics, Humans, Induced Pluripotent Stem Cells immunology, Induced Pluripotent Stem Cells metabolism, Plasmids genetics, Tissue Donors, Cell Culture Techniques methods, Induced Pluripotent Stem Cells cytology
- Abstract
We report a simple method, using p53 suppression and nontransforming L-Myc, to generate human induced pluripotent stem cells (iPSCs) with episomal plasmid vectors. We generated human iPSCs from multiple donors, including two putative human leukocyte antigen (HLA)-homozygous donors who match ∼20% of the Japanese population at major HLA loci; most iPSCs are integrated transgene-free. This method may provide iPSCs suitable for autologous and allologous stem-cell therapy in the future. more...
- Published
- 2011
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11. Generation of mouse-induced pluripotent stem cells with plasmid vectors.
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Okita K, Hong H, Takahashi K, and Yamanaka S
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- Animals, Base Sequence, DNA Primers genetics, Embryonic Stem Cells cytology, Embryonic Stem Cells metabolism, Fibroblasts cytology, Genes, myc, Humans, Kruppel-Like Factor 4, Kruppel-Like Transcription Factors genetics, Mice, Octamer Transcription Factor-3 genetics, Pluripotent Stem Cells metabolism, SOXB1 Transcription Factors genetics, Transfection, Cell Dedifferentiation genetics, Genetic Vectors, Plasmids genetics, Pluripotent Stem Cells cytology
- Abstract
Reprogramming of somatic cells into pluripotent stem cells has been reported by introducing a combination of several transcription factors (Oct3/4, Sox2, Klf4 and c-Myc). The induced pluripotent stem (iPS) cells from patient's somatic cells could be a useful source for drug discovery and cell transplantation therapies. However, to date, most iPS cells were made using viral vectors, such as retroviruses and lentiviruses. Here we describe an alternative method to generate iPS cells from mouse embryonic fibroblasts (MEFs) by continual transfection of plasmid vectors. This protocol takes around 2 months to complete, from MEF isolation to iPS cell establishment. Although the reprogramming efficiency of this protocol is still low, the established iPS cells are most likely free from plasmid integration. This virus-free technique reduces the safety concern for iPS cell generation and application, and provides a source of cells for the investigation of the mechanisms underlying reprogramming and pluripotency. more...
- Published
- 2010
- Full Text
- View/download PDF
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