Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells

Santosh Mathapati; Richard Siller; Agata A R Impellizzeri; Max Lycke; Karianne Vegheim; Runar Almaas; Gareth J Sullivan

doi:10.1002/cpsc.13

Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells

Santosh Mathapati, Richard Siller, Agata A R Impellizzeri, Max Lycke, Karianne Vegheim, Runar Almaas, Gareth J Sullivan

School of Medicine

Research output: Contribution to journal › Article › peer-review

34 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1G.6.1-1G.6.18
Journal	Current protocols in stem cell biology
Volume	38
DOIs	https://doi.org/10.1002/cpsc.13
Publication status	Published - 17 Aug 2016

Keywords

Cell Differentiation/drug effects
Endoderm/cytology
Feeder Cells/cytology
Hepatocytes/cytology
Humans
Pluripotent Stem Cells/cytology
Small Molecule Libraries/pharmacology
Tissue Culture Techniques/methods

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/cpsc.13

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title = "Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells",

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. {\textcopyright} 2016 by John Wiley & Sons, Inc.",

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author = "Santosh Mathapati and Richard Siller and Impellizzeri, {Agata A R} and Max Lycke and Karianne Vegheim and Runar Almaas and Sullivan, {Gareth J}",

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doi = "10.1002/cpsc.13",

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AU - Mathapati, Santosh

AU - Siller, Richard

AU - Impellizzeri, Agata A R

AU - Lycke, Max

AU - Vegheim, Karianne

AU - Almaas, Runar

AU - Sullivan, Gareth J

PY - 2016/8/17

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N2 - 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.

AB - 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.

KW - Cell Differentiation/drug effects

KW - Endoderm/cytology

KW - Feeder Cells/cytology

KW - Hepatocytes/cytology

KW - Humans

KW - Pluripotent Stem Cells/cytology

KW - Small Molecule Libraries/pharmacology

KW - Tissue Culture Techniques/methods

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DO - 10.1002/cpsc.13

M3 - Article

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SP - 1G.6.1-1G.6.18

JO - Current protocols in stem cell biology

JF - Current protocols in stem cell biology

ER -

Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells

Abstract

Keywords

UN SDGs

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