# Comparative analysis of small molecule and growth factor-derived human induced pluripotent stem cell-derived hepatocyte-like cells

## TL;DR

This study compares two methods for generating liver-like cells from stem cells and finds that one method produces more mature liver-like cells suitable for metabolism and infection studies.

## Contribution

The study provides a comparative analysis of growth factor and small molecule protocols for generating hepatocyte-like cells from human iPSCs.

## Key findings

- Growth factor-derived HLCs show mature hepatocyte morphology and gene/protein expression.
- Small molecule-derived HLCs resemble liver tumor-derived cells with a dedifferentiated phenotype.
- Growth factor-derived HLCs are better suited for metabolism and viral infection studies.

## Abstract

The growth factor and small molecule protocol are the two primary approaches for generating human induced pluripotent stem cell-derived hepatocyte-like cells (iPSC-HLCs). We compared the efficacy of the growth factor and small molecule protocols across fifteen different human iPSC lines. Morphological assessment, relative quantification of gene expression, protein expression and proteomic studies were carried out. HLCs derived from the growth factor protocol displayed mature hepatocyte morphological features including a raised, polygonal shape with well-defined refractile borders, granular cytoplasm with lipid droplets and/or vacuoles with multiple spherical nuclei or a large centrally located nucleus; significantly elevated hepatocyte gene and protein expression including AFP, HNF4A, ALBUMIN, and proteomic and metabolic features that are more aligned with a mature phenotype. HLCs derived from the small molecule protocol showed a dedifferentiated, proliferative phenotype that is more akin to liver tumor-derived cell lines. These experimental results suggest that HLCs derived from growth factors are better suited for studies of metabolism, biotransformation, and viral infection.