Study summary · research use only
Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
This methods article (in vitro, human pluripotent stem cells) describes a small-molecule-driven protocol for differentiating human pluripotent stem cells (hPSCs) into hepatocyte-like cells (HLCs) without added growth factors. hPSCs are directed through a primitive-streak route to definitive endoderm (DE) using the small molecule CHIR99021, a Wnt agonist, in place of Wnt3A and activin A. The DE is then differentiated into hepatoblast-like cells using dimethyl sulfoxide, and the hepatoblasts are further differentiated into HLCs using N-hexanoic-Tyr, Ile-6 aminohexanoic amide (Dihexa, a hepatocyte growth factor agonist) and dexamethasone. The authors describe the protocol as an efficient and reproducible procedure for hPSC-to-HLC differentiation using small molecules.
Abstract
Hepatocyte-like cells (HLCs) generated in vitro from human pluripotent stem cells (hPSCs) provide an invaluable resource for basic research, regenerative medicine, drug screening, toxicology, and modeling of liver disease and development. This unit describes a small-molecule-driven protocol for in vitro differentiation of hPSCs into HLCs without the use of growth factors. hPSCs are coaxed through a developmentally relevant route via the primitive streak to definitive endoderm (DE) using the small molecule CHIR99021 (a Wnt agonist), replacing the conventional growth factors Wnt3A and activin A. The small-molecule-derived DE is then differentiated to hepatoblast-like cells in the presence of dimethyl sulfoxide. The resulting hepatoblasts are then differentiated to HLCs with N-hexanoic-Tyr, Ile-6 aminohexanoic amide (Dihexa, a hepatocyte growth factor agonist) and dexamethasone. The protocol provides an efficient and reproducible procedure for differentiation of hPSCs into HLCs utilizing small molecules. © 2016 by John Wiley & Sons, Inc.
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