Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells

Rajagopal N Aravalli,1 Daniel P Collins,2 Joel H Hapke,2 Andrew T Crane,3 Clifford J Steer4,5 1Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, MN 55455, USA; 2Cytomedical Design Group LLC, St. Paul, MN 55127, USA; 3Department of Neurosurgery, University of Mi...

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Autores principales: Aravalli RN, Collins DP, Hapke JH, Crane AT, Steer CJ
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Publicado: Dove Medical Press 2020
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spelling oai:doaj.org-article:e71c8ce8956e4de0aa96d3ebe9257b042021-12-02T03:32:52ZHepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells1179-1535https://doaj.org/article/e71c8ce8956e4de0aa96d3ebe9257b042020-02-01T00:00:00Zhttps://www.dovepress.com/hepatic-differentiation-of-marmoset-embryonic-stem-cells-and-functiona-peer-reviewed-article-HMERhttps://doaj.org/toc/1179-1535Rajagopal N Aravalli,1 Daniel P Collins,2 Joel H Hapke,2 Andrew T Crane,3 Clifford J Steer4,5 1Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, MN 55455, USA; 2Cytomedical Design Group LLC, St. Paul, MN 55127, USA; 3Department of Neurosurgery, University of Minnesota Medical School, Minneapolis, MN 55455, USA; 4Department of Medicine, University of Minnesota Medical School, Minneapolis, MN 55455, USA; 5Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USACorrespondence: Rajagopal N AravalliDepartment of Electrical and Computer Engineering, University of Minnesota, 200 Union Street SE, Minneapolis, MN 55455, USAEmail aravalli@umn.eduClifford J SteerDepartment of Medicine, University of Minnesota, 420 Delaware Street SE, Minneapolis, MN 55455, USAEmail steer001@umn.eduBackground: Primary human hepatocytes (PHHs) are the ideal candidates for studying critical liver functions such as drug metabolism and toxicity. However, as they are isolated from discarded livers that are unsuitable for transplantation, they possess limited expansion ability in vitro and their enzymatic functions deteriorate rapidly because they are often of poor quality. Therefore, there is a compelling reason to find reliable alternative sources of hepatocytes.Methods: In this study, we report on efficient and robust differentiation of embryonic stem cells (ESC) from the common marmoset Callithrix jacchus into functional hepatocyte-like cells (HLC) using a simple, and reproducible three-step procedure. ESC-derived HLCs were examined by morphological analysis and tested for their expression of hepatocyte-specific markers using a combination of immunohistochemistry, RT-PCR, and biochemical assays. Primary human hepatocytes were used as controls.Results: ESC-derived HLCs expressed each of the hepatocyte-specific markers tested, including albumin; α-fetoprotein; asialoglycoprotein receptor 1; α-1 antitrypsin; hepatocyte nuclear factors 1α and 4; cytokeratin 18; hepatocyte growth factor receptor; transferrin; tyrosine aminotransferase; alkaline phosphatase; c-reactive protein; cytochrome P450 enzymes CYP1A2, CYP2E1 and CYP3A4; and coagulation factors FVII and FIX. They were functionally competent as demonstrated by biochemical assays in addition to producing urea.Conclusion: Our data strongly suggest that marmoset HLCs possess characteristics similar to those of PHHs. They could, therefore, be invaluable for studies on drug metabolism and cell transplantation therapy for a variety of liver disorders. Because of the similarities in the anatomical and physiological features of the common marmoset to that of humans, Callithrix jacchus is an appropriate animal model to study human disease conditions and cellular functions.Keywords: Callithrix jacchus, marmoset, hepatocyte, embryonic stem cell, differentiation, liverAravalli RNCollins DPHapke JHCrane ATSteer CJDove Medical Pressarticlecallithrix jacchusmarmosethepatocyteembryonic stem celldifferentiationliverDiseases of the digestive system. GastroenterologyRC799-869ENHepatic Medicine: Evidence and Research, Vol Volume 12, Pp 15-27 (2020)
institution DOAJ
collection DOAJ
language EN
topic callithrix jacchus
marmoset
hepatocyte
embryonic stem cell
differentiation
liver
Diseases of the digestive system. Gastroenterology
RC799-869
spellingShingle callithrix jacchus
marmoset
hepatocyte
embryonic stem cell
differentiation
liver
Diseases of the digestive system. Gastroenterology
RC799-869
Aravalli RN
Collins DP
Hapke JH
Crane AT
Steer CJ
Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
description Rajagopal N Aravalli,1 Daniel P Collins,2 Joel H Hapke,2 Andrew T Crane,3 Clifford J Steer4,5 1Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, MN 55455, USA; 2Cytomedical Design Group LLC, St. Paul, MN 55127, USA; 3Department of Neurosurgery, University of Minnesota Medical School, Minneapolis, MN 55455, USA; 4Department of Medicine, University of Minnesota Medical School, Minneapolis, MN 55455, USA; 5Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USACorrespondence: Rajagopal N AravalliDepartment of Electrical and Computer Engineering, University of Minnesota, 200 Union Street SE, Minneapolis, MN 55455, USAEmail aravalli@umn.eduClifford J SteerDepartment of Medicine, University of Minnesota, 420 Delaware Street SE, Minneapolis, MN 55455, USAEmail steer001@umn.eduBackground: Primary human hepatocytes (PHHs) are the ideal candidates for studying critical liver functions such as drug metabolism and toxicity. However, as they are isolated from discarded livers that are unsuitable for transplantation, they possess limited expansion ability in vitro and their enzymatic functions deteriorate rapidly because they are often of poor quality. Therefore, there is a compelling reason to find reliable alternative sources of hepatocytes.Methods: In this study, we report on efficient and robust differentiation of embryonic stem cells (ESC) from the common marmoset Callithrix jacchus into functional hepatocyte-like cells (HLC) using a simple, and reproducible three-step procedure. ESC-derived HLCs were examined by morphological analysis and tested for their expression of hepatocyte-specific markers using a combination of immunohistochemistry, RT-PCR, and biochemical assays. Primary human hepatocytes were used as controls.Results: ESC-derived HLCs expressed each of the hepatocyte-specific markers tested, including albumin; α-fetoprotein; asialoglycoprotein receptor 1; α-1 antitrypsin; hepatocyte nuclear factors 1α and 4; cytokeratin 18; hepatocyte growth factor receptor; transferrin; tyrosine aminotransferase; alkaline phosphatase; c-reactive protein; cytochrome P450 enzymes CYP1A2, CYP2E1 and CYP3A4; and coagulation factors FVII and FIX. They were functionally competent as demonstrated by biochemical assays in addition to producing urea.Conclusion: Our data strongly suggest that marmoset HLCs possess characteristics similar to those of PHHs. They could, therefore, be invaluable for studies on drug metabolism and cell transplantation therapy for a variety of liver disorders. Because of the similarities in the anatomical and physiological features of the common marmoset to that of humans, Callithrix jacchus is an appropriate animal model to study human disease conditions and cellular functions.Keywords: Callithrix jacchus, marmoset, hepatocyte, embryonic stem cell, differentiation, liver
format article
author Aravalli RN
Collins DP
Hapke JH
Crane AT
Steer CJ
author_facet Aravalli RN
Collins DP
Hapke JH
Crane AT
Steer CJ
author_sort Aravalli RN
title Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
title_short Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
title_full Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
title_fullStr Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
title_full_unstemmed Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells
title_sort hepatic differentiation of marmoset embryonic stem cells and functional characterization of esc-derived hepatocyte-like cells
publisher Dove Medical Press
publishDate 2020
url https://doaj.org/article/e71c8ce8956e4de0aa96d3ebe9257b04
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