Methods Mol Biol. 2019;1994:41-53. doi: 10.1007/978-1-4939-9477-9_4.

Human Induced Pluripotent Stem Cell-Derived Definitive Endoderm Bulk Culture and Hepatic Differentiation.

Palakkan AA¹, Nanda J¹, Ross JA².

Abstract

We have developed a method to bulk culture definitive endoderm cells generated from human iPSCs which can be stored and differentiated to hepatocytes. Human iPSC-derived definitive endoderm cells were sorted based on the expression of CXCR4. The sorted cells were able to proliferate for extended periods and can be cryopreserved. The definitive endoderm cells were subsequently utilized to generate functional hepatocytes expressing albumin and α-fetoprotein in different multiwell formats. This provides a method to reliably produce more consistent hepatocytes in greater quantities and has enabled the development of high-throughput screening strategies.