iPSC-derived hepatocytes generated from NASH donors provide a valuable platform for disease modeling and drug discovery
| Autor: | Deepika Rajesh, Makiko Ohshima, Sarah Burton, Christie Munn, Madelyn E Goedland, Igor Gurevich, Katherine Czysz |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular Cirrhosis QH301-705.5 Science Cellular differentiation medicine.medical_treatment Induced Pluripotent Stem Cells Cell Culture Techniques Biology Liver transplantation digestive system General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine In vivo Non-alcoholic Fatty Liver Disease Fatty liver disease Drug Discovery medicine Humans Hepatocyte Biology (General) Cells Cultured 030304 developmental biology Cryopreservation 0303 health sciences Fatty liver Endoderm NASH nutritional and metabolic diseases Cell Differentiation medicine.disease Flow Cytometry digestive system diseases medicine.anatomical_structure Stem cell differentiation Cancer research Hepatocytes 030211 gastroenterology & hepatology Steatohepatitis Co-culture General Agricultural and Biological Sciences Liver cancer Biomarkers Research Article |
| Zdroj: | Biology Open article-version (VoR) Version of Record Biology Open, Vol 9, Iss 12 (2020) |
| ISSN: | 2046-6390 |
| Popis: | Non-alcoholic fatty liver disease (NAFLD) affects 30–40% of adults and 10% of children in the US. About 20% of people with NAFLD develop non-alcoholic steatohepatitis (NASH), which may lead to cirrhosis and liver cancer, and is projected to be a leading cause of liver transplantation in the near future. Human induced pluripotent stem cells (iPSC) from NASH patients are useful for generating a large number of hepatocytes for NASH modeling applications and identification of potential drug targets. We developed a novel defined in vitro differentiation process to generate cryopreservable hepatocytes using an iPSC panel of NASH donors and apparently healthy normal (AHN) controls. iPSC-derived hepatocytes displayed stage specific phenotypic markers, hepatocyte morphology, with bile canaliculi. Importantly, both fresh and cryopreserved definitive endoderm and hepatoblasts successfully differentiated to pure and functional hepatocytes with increased CYP3A4 activity in response to rifampicin and lipid accumulation upon fatty acid (FA) treatment. End-stage hepatocytes integrated into three-dimensional (3D) liver organoids and demonstrated increased levels of albumin secretion compared to aggregates consisting of hepatocytes alone. End-stage hepatocytes derived from NASH donors demonstrated spontaneous lipidosis without FA supplementation, recapitulating a feature of NASH hepatocytes in vivo. Cryopreserved hepatocytes generated by this protocol across multiple donors will provide a critical cell source to facilitate the fundamental understanding of NAFLD/NASH biology and potential high throughput screening applications for preclinical evaluation of therapeutic targets. Summary: We describe the development and validation of a hepatocyte differentiation protocol using iPSCs from a panel of NASH donors and healthy controls. |
| Databáze: | OpenAIRE |
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