Protocol for the generation of pancreatic and hepatic progenitors from human pluripotent stem cells for gene regulatory assays

Lay Shuen Tan; Resilind Su Ern Chew; Natasha Hui Jin Ng; Adrian Kee Keong Teo

doi:10.1016/j.xpro.2021.100471

Protocol for the generation of pancreatic and hepatic progenitors from human pluripotent stem cells for gene regulatory assays

STAR Protoc. 2021 Apr 19;2(2):100471. doi: 10.1016/j.xpro.2021.100471. eCollection 2021 Jun 18.

Authors

Lay Shuen Tan¹, Resilind Su Ern Chew¹, Natasha Hui Jin Ng¹, Adrian Kee Keong Teo^{1

2}

Affiliations

¹ Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology (IMCB), A∗STAR, Singapore 138673, Singapore.
² Department of Biochemistry and Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596, Singapore.

Abstract

This protocol describes the detailed procedures for utilizing human pluripotent stem cells (hPSCs) for pancreatic progenitor and hepatic differentiation, followed by the application of hPSC-derived cells in a luciferase reporter-based assay to study gene regulation. The generated hPSC-derived cells have been shown to achieve morphologies and gene expression profiles specific to their differentiated cell types, and subsequent luciferase assay has been shown to effectively elucidate the role of disease-relevant gene variants. Therefore, this protocol provides a valuable approach for pancreatic and liver disease modeling. For complete details on the use and execution of this protocol, please refer to Ng et al. (2019).

Keywords: Cell Differentiation; Cell culture; Cell-based Assays; Gene Expression; Molecular Biology; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation*
Cells, Cultured
Gene Expression Profiling*
Gene Expression Regulation*
Humans
Liver / cytology
Liver / metabolism*
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism*