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3. Phenotypical characterization of 6–21-week gestational age human dermis and epidermal cell isolation methods for in vitro studies on epidermal progenitors

Author

Johnen, Christa, Chinnici, Cinzia, Triolo, Fabio, Plettig, Jörn, Bräutigam, Kirsten, Amico, Giandomenico, Young, Matthew, Over, Patrick, Esteban-Vives, Roger, Schmelzer, Eva, Conaldi, Pier-Giulio, Turner, Morris, Thompson, Robert, Zeilinger, Katrin, Rubin, Peter, Vizzini, Giovanni, Gridelli, Bruno, and Gerlach, Jörg C.

Published
2013
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7. Hedgehog signaling maintains resident hepatic progenitors throughout life

Author

Sicklick, Jason K., Li, Yin-Xiong, Melhem, Alaa, Schmelzer, Eva, Zdanowicz, Marzena, Huang, Jiawen, Caballero, Montserrat, Fair, Jeff H., Ludlow, John W., McClelland, Randall E., Reid, Lola M., and Diehl, Anna Mae

Subjects
Genetic transcription -- Research, Hedgehogs -- Health aspects, Hedgehogs -- Genetic aspects, Biological sciences
Abstract

Hedgehog signaling through its receptor, Patched, activates transcription of genes, including Patched, that regulate the fate of various progenitors. Although Hedgehog signaling is required for endodermal commitment and hepatogenesis, the possibility that it regulates liver turnover in adults had not been considered because mature liver epithelial cells lack Hedgehog signaling. Herein, we show that this pathway is essential throughout life for maintaining hepatic progenitors. Patched-expressing cells have been identified among endodermally lineage-restricted, murine embryonic stem cells as well as in livers of fetal and adult Ptc-lacZ mice. An adult-derived, murine hepatic progenitor cell line expresses Patched, and Hedgehog-responsive cells exist in stein cell compartments of fetal and adult human livers. In both species, manipulation of Hedgehog activity influences hepatic progenitor cell survival. Therefore, Hedgehog signaling is conserved in hepatic progenitors from fetal development through adulthood and may be a new therapeutic target in patients with liver damage. Indian hedgehog; liver; Patched; progenitor cell; Sonic hedgehog

Published
2006

8. Conditioned Medium from Human Amnion-Derived Mesenchymal Stromal/Stem Cells Attenuating the Effects of Cold Ischemia-Reperfusion Injury in an In Vitro Model Using Human Alveolar Epithelial Cells

Author

Miceli, Vitale, primary, Bertani, Alessandro, additional, Chinnici, Cinzia Maria, additional, Bulati, Matteo, additional, Pampalone, Mariangela, additional, Amico, Giandomenico, additional, Carcione, Claudia, additional, Schmelzer, Eva, additional, Gerlach, Jörg C., additional, and Conaldi, Pier Giulio, additional

Published
2021
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12. Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance

Author

Wauthier, Eliane, primary, Schmelzer, Eva, additional, Turner, William, additional, Zhang, Lili, additional, LeCluyse, Ed, additional, Ruiz, Joseph, additional, Turner, Rachael, additional, Furth, M.E., additional, Kubota, Hiroshi, additional, Lozoya, Oswaldo, additional, Barbier, Claire, additional, McClelland, Randall, additional, Yao, Hsin‐lei, additional, Moss, Nicholas, additional, Bruce, Andrew, additional, Ludlow, John, additional, and Reid, L.M., additional

Published
2008
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17. Interwoven Four-Compartment Capillary Membrane Technology for Three-Dimensional Perfusion with Decentralized Mass Exchange to Scale Up Embryonic Stem Cell Culture

Author

Gerlach, Jörg C., Lübberstedt, Marc, Edsbagge, Josefina, Ring, Alexander, Hout, Mariah, Baun, Matt, Rossberg, Ingrid, Knöspel, Fanny, Peters, Grant, Eckert, Klaus, Wulf-Goldenberg, Annika, Björquist, Petter, Stachelscheid, Harald, Urbaniak, Thomas, Schatten, Gerald, Miki, Toshio, Schmelzer, Eva, and Zeilinger, Katrin

Published
2010
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21. The degradation behavior of calcium‐rich hydroxyapatite foams in vitro.

Author

Zhang, Qinghao, Wang, Wenfu, Schmelzer, Eva, Gerlach, Jörg, Liu, Changsheng, and Nettleship, Ian

Abstract

Hydroxyapatite (HA) is a well‐known regenerative biomaterial. However, the slow degradation rate of HA is still an obstacle in clinical applications. In this study, we concentrated on investigating the degradation behavior of the calcium‐rich HA foams, which had a demonstrated effect on blood differentiation in previous studies. The HA foams were processed by an emulsion method and were infiltrated with calcium nitrate to create a calcium carbonate second phase, heterogeneously distributed on and under the surface of the foam. During the 28‐day solubility test, the calcium carbonate phase contributed to enhanced Ca2+ ion release into the saline compared to phase pure HA foams. Both types of foams were biocompatible as demonstrated by human endothelial cell culture on their surface. The release of calcium ions, the degradation behavior, and the endothelial cell differentiation behavior suggest this biphasic ceramic is a candidate for bone marrow in vitro culture and a possible bone substitute material. [ABSTRACT FROM AUTHOR]

Published
2021
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30. Transplantation of hepatocytes from genetically engineered pigs into baboons

Author

Iwase, Hayato, primary, Liu, Hong, additional, Schmelzer, Eva, additional, Ezzelarab, Mohamed, additional, Wijkstrom, Martin, additional, Hara, Hidetaka, additional, Lee, Whayoung, additional, Singh, Jagjit, additional, Long, Cassandra, additional, Lagasse, Eric, additional, Gerlach, Jörg C., additional, Cooper, David K. C., additional, and Gridelli, Bruno, additional

Published
2017
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33. Epithelial cell adhesion molecule fragments and signaling in primary human liver cells.

Author

Gerlach, Jörg C., Foka, Hubert G., Thompson, Robert L., Gridelli, Bruno, and Schmelzer, Eva

Subjects
EPITHELIAL cells, CELL adhesion molecules, GLYCOPROTEIN synthesis, MEMBRANE proteins, CELL physiology
Abstract

Epithelial Cell Adhesion Molecule (EpCAM), or CD326, is a trans‐membrane glycoprotein expressed by multiple normal epithelia as well as carcinoma. Human hepatic stem cells and bile duct epithelium of the liver are EpCAM positive. In tumor cell lines, its intracellular domain can be released after cleavage of the extracellular domain. Within the cell nucleus, it induces cell proliferation, but cleavage depends on cell contact. Fragments of various lengths have been described in tumor cells. Despite its described important role in proliferation in tumor cells, there is not much known about the expression and role of EpCAM fragments in primary human liver cells. Here, we demonstrate that EpCAM protein fragments and function are considerable different between tumor cells, normal fetal and adult liver cells. Contrary to previously reported findings in tumor cells, gene knockdown or treatment with an inhibitor of the cleavage enzyme ADAM17 (TACE) rather increased cell numbers in primary human fetal liver‐derived EpCAM‐positive cells. EpCAM fragment sizes were not affected by treatment with inhibitor. Knockdown of EPCAM gene expression by siRNA in sorted cells did not significantly affect proliferation‐associated genes or cell numbers. The intracellular domain could not be detected within cell nuclei of fetal and adult liver cells. In conclusion, signaling through the intracellular domain of EpCAM appears to be a mechanism that induces proliferation specifically in tumorigenic cells but not in normal primary EpCAM‐positive liver cells. [ABSTRACT FROM AUTHOR]

Published
2018
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35. Phases I-II Matched Case-Control Study of Human Fetal Liver Cell Transplantation for Treatment of Chronic Liver Disease

Author

Pietrosi, Giada, primary, Vizzini, Giovanni, additional, Gerlach, Jorg, additional, Chinnici, Cinzia, additional, Luca, Angelo, additional, Amico, Giandomenico, additional, D'amato, Monica, additional, Conaldi, Pier Giulio, additional, Petri, Sergio Li, additional, Spada, Marco, additional, Tuzzolino, Fabio, additional, Alio, Luigi, additional, Schmelzer, Eva, additional, and Gridelli, Bruno, additional

Published
2015
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40. Ex Vivo Conditions for Self-Replication of Human Hepatic Stem Cells

Author

Barbier, Claire, Reid, Lola M., Melhem, Alaa, Wauthier, Eliane, Zhang, Lili, McClelland, Randall, and Schmelzer, Eva

Abstract

Human hepatic stem cells (hHpSCs), identifiable by a unique antigenic profile, have been isolated from human livers and established ex vivo under expansion conditions permissive for self-replication. The conditions consist of a substratum of type III collagen, ideally on Transwell inserts, and Kubota's medium, a serum-free medium developed for hepatic progenitors. Under these conditions the cells demonstrated a doubling time of ∼24 h, generating at least a 16-fold increase in cell number within 7–10 days; were stable at confluence for up to 2 weeks; could be passaged, if on type III collagen, to initiate colonies that went through log-phase growth and saturation density kinetics; and expressed telomerase, indicative of regenerative capacity. The hHpSC colonies remained morphologically and phenotypically stable throughout expressing epithelial cell adhesion molecule, neural cell adhesion molecule, albumin, cytokeratins 8, 18, and 19, but not α-fetoprotein, or intercellular adhesion molecule-1 (ICAM-1). Those maintained under self-replication conditions for more than a month were transplanted and found to engraft in the livers of SCID/nod mice yielding human liver tissue expressing adult liver–specific proteins. The conditions for self-replication should offer ideal culture conditions for generating large numbers of hHpSCs for use in commercial and clinical programs.

Published
2008
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42. Optimierung der Kultur und Charakterisierung primärer embryonaler Hepatozyten der Ratte

Author

Schmelzer, Eva and Deckwer, Wolf Dieter

Subjects
ddc:57, doctoral thesis, ddc:570, ddc:5
Abstract

In dieser Arbeit wurde in vitro die Proliferation und Differenzierung embryonaler Hepatozyten untersucht. Im Rattenmodell wurden daher die Kulturbedingungen der Hepatozyten variiert und optimiert. Mit Hilfe von Analysen sowohl auf genomischer als auch funktioneller Ebene wurde eine umfassende Charakterisierung der Hepatozyten in Kultur durchgeführt. Untersucht wurde der Einfluß verschiedener Matrizes (Collagen I, IV, Matrigel, Laminin), feeder-Zellen (3T3, MRC-5, primäre Fibroblasten) und Wachstumsfaktoren einzeln oder in Kombination (Thrombopoietin [TPO], Hepatocyte-, Epidermal- und Transforming Growth Factor) auf die Proliferation und Differenzierung der embryonalen Hepatozyten. Die Proliferation und Differenzierung embryonaler Hepatozyten in Kultur werden eindeutig von der Art der verwendeten Beschichtung bzw. feeder-Zellen beeinflußt. Maximale, über 30fache Zellvermehrung, wurde nach 12 Tagen in Kultur auf 3T3 feeder-Zellen mit Zusatz von TPO erreicht. Es konnte erstmalig ein proliferativer Effekt von TPO auf Hepatozyten in Kultur gezeigt werden. Die auf 3T3 feeder-Zellen mit TPO kultivierten Zellen wiesen ein RNA Expressionsprofil ähnlich dem adulter Hepatozyten auf. Zusätzlich wurden Differenzierungsmarker nachgewiesen: Es konnte Albuminsekretion über einen Untersuchungszeitraum von 30 Tagen gemessen werden; durch Immuncytochemie wurden Albumin, GST und verschiedene Cytochrome detektiert; es konnte Cytochrom-P450 Aktivität nachgewiesen werden., The aim of the present dissertation was to investigate in vitro proliferation and differentiation of embryonic rat hepatocytes. Therefore, culture conditions of hepatocytes were varied and optimised. A widespread characterisation of embryonic hepatocytes in culture was realised by using genomic as well as functional methods of analysis. The influence of various matrices (collagen I, IV, matrigel, laminin), feeder cells (3T3, MRC-5, primary fibroblasts) and growth factors (thrombopoietin [TPO], hepatocyte growth factor, epidermal growth factor and transforming growth factor alpha) alone or in combination on proliferation and differentiation of these embryonic hepatocytes was studied. Proliferation and differentiation of embryonic hepatocytes in culture is clearly affected by type of coating and feeder cell, respectively. After 12 days in culture maximal increase in cell number (more than 30 fold) was observed using 3T3 as feeder cells in combination with TPO. For the first time a proliferative effect of TPO on hepatocytes could be demonstrated. The RNA profile of expression of hepatocytes cultured on 3T3 feeder cells supplemented with TPO bear resemblance to that of adult hepatocytes. In addition, markers of differentiation could be verified. Secretion of albumin could be measured during an observation period of 30 days. Immuncytochemistry revealed expression of albumin, GST, and various cytochromes. Cytochrome P450 activities could be detected for several days in culture.

Published
2002

43. Hepatic stem cells and hepatoblasts : identification, isolation, and ex vivo maintenance

Author

Wauthier, Eliane, Schmelzer, Eva, Turner, William, Zhang, Lili, LeCluyse, E, Ruiz, Joseph, Turner, Rachael, Furth, Mark E., Kubota, Hiroshi, Lozoya, Oswaldo, Barbier, Claire, McClelland, Randall E, Yao, Hsin Lei, Moss, Nicholas, Bruce, Andrew, Ludlow, John, Reid, Lola M., Wauthier, Eliane, Schmelzer, Eva, Turner, William, Zhang, Lili, LeCluyse, E, Ruiz, Joseph, Turner, Rachael, Furth, Mark E., Kubota, Hiroshi, Lozoya, Oswaldo, Barbier, Claire, McClelland, Randall E, Yao, Hsin Lei, Moss, Nicholas, Bruce, Andrew, Ludlow, John, and Reid, Lola M.

Published
2008

45. Compartmental Hollow Fiber Capillary Membrane–Based Bioreactor Technology for In Vitro Studies on Red Blood Cell Lineage Direction of Hematopoietic Stem Cells

Author

Housler, Greggory J., primary, Miki, Toshio, additional, Schmelzer, Eva, additional, Pekor, Christopher, additional, Zhang, Xiaokui, additional, Kang, Lin, additional, Voskinarian-Berse, Vanessa, additional, Abbot, Stewart, additional, Zeilinger, Katrin, additional, and Gerlach, Jörg C., additional

Published
2012
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