Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology (IMCB), A∗STAR, Singapore 138673, Singapore.
Department of Biochemistry and Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596, Singapore.
STAR Protoc. 2021 Apr 19;2(2):100471. doi: 10.1016/j.xpro.2021.100471. eCollection 2021 Jun 18.
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).
本方案描述了利用人类多能干细胞（hPSC）进行胰腺祖细胞和肝分化的详细步骤，随后将 hPSC 衍生细胞应用于基于荧光素酶报告基因的检测方法中，以研究基因调控。已经证明生成的 hPSC 衍生细胞具有与其分化细胞类型特异性的形态和基因表达谱，并且随后的荧光素酶检测已被证明能够有效地阐明与疾病相关的基因变异体的作用。因此，本方案为胰腺和肝脏疾病建模提供了一种有价值的方法。有关本方案使用和执行的完整详细信息，请参见 Ng 等人（2019 年）。
