Generation of SST-P2A-mCherry reporter human embryonic stem cell line using the CRISPR/Cas9 system (WAe001-A-2C)

Tongran Zhang; Feng Zhang; Nannan Wang; Tao Xu; Lingqiang Zhu; Lihua Chen; Huisheng Liu

doi:10.1016/j.scr.2024.103397

Generation of SST-P2A-mCherry reporter human embryonic stem cell line using the CRISPR/Cas9 system (WAe001-A-2C)

Stem Cell Res. 2024 Jun:77:103397. doi: 10.1016/j.scr.2024.103397. Epub 2024 Mar 21.

Authors

Tongran Zhang¹, Feng Zhang², Nannan Wang³, Tao Xu⁴, Lingqiang Zhu⁵, Lihua Chen⁶, Huisheng Liu⁷

Affiliations

¹ Department of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430074, China; Guangzhou Laboratory, Guangzhou 510006, China.
² Guangzhou Laboratory, Guangzhou 510006, China; The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510006, China.
³ Guangzhou Laboratory, Guangzhou 510006, China; College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
⁴ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou Laboratory, Guangzhou 510006, China.
⁵ Department of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430074, China. Electronic address: [email protected].
⁶ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou Laboratory, Guangzhou 510006, China. Electronic address: [email protected].
⁷ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou Laboratory, Guangzhou 510006, China; School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou 510006, China; College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China. Electronic address: [email protected].

PMID: 38547668
DOI: 10.1016/j.scr.2024.103397

Abstract

Somatostatin (SST)-producing pancreatic delta-cells play an important role in maintaining the balance of insulin and glucagon secretion within the islets. This study aimed to generate a human embryonic stem cell (hESC) line with a SST-P2A-mCherry reporter using CRISPR/Cas9 system. The SST-P2A-mCherry reporter cell line was shown to maintain typical pluripotent characteristics and able to be induced into SST-producing pancreatic delta-cells. The generation of the cell line would provide useful platform for the characterization of stem cell-derived delta-cells, discovery of delta-cell surface markers and investigation of paracrine mechanisms, which will ultimately promote the drug discovery and cell therapy of diabetes mellitus.

MeSH terms

CRISPR-Cas Systems*
Cell Differentiation
Cell Line
Genes, Reporter
Human Embryonic Stem Cells* / cytology
Human Embryonic Stem Cells* / metabolism
Humans
Somatostatin / metabolism

Substances

Somatostatin