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METHODS article
Front. Cell Dev. Biol.
Sec. Stem Cell Research
Volume 12 - 2024 |
doi: 10.3389/fcell.2024.1490040
A Differentiation Protocol for Generating Pancreatic Delta Cells from Human Pluripotent Stem Cells
Provisionally accepted- 1 Department of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
- 2 Guangzhou National Laboratory, Guangzhou, China
In this protocol, we detail a seven-stage differentiation methodology for generating pancreatic delta cells (SC-delta cells) from human pluripotent stem cells (hPSCs). In the first step, definitive endoderm is generated by activin A and CHIR99021, followed by induction of primitive gut tube and posterior foregut by treatment with FGF7, SANT1, LDN193189, PdBU and retinoic acid (RA). The subsequent endocrine generation and directed SC-delta cell induction is achieved by a combined treatment of the FGF7 with FGF2 during stage 4 and 5, together with RA, XXI, ALK5 inhibitor II, SANT1, Betacellulin and LDN193189. The planar cultivation is converted to a suspended system after stage 5, allowing cells to aggregate into delta cell-containing spheroids. The differentiation takes approximately 4-5 weeks for delta cell generation and an additional 1-2 weeks for cell expansion and evaluation. We believe that this amenable and simplified protocol can provide a stable source of SC-delta cells from efficient differentiation, facilitating further investigation of the physiological role of delta cells as well as refinement of islet cell therapeutic strategies.
Keywords: hPSCs, stem cell differentiation, pancreatic delta cells, Fgf, islet organoids
Received: 02 Sep 2024; Accepted: 07 Oct 2024.
Copyright: © 2024 Zhang, Wang, Liao, Chen, Meng, Lin, Xu, Chen, Zhu and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Nannan Wang, Guangzhou National Laboratory, Guangzhou, China
Jingyi Chen, Guangzhou National Laboratory, Guangzhou, China
Hao Meng, Guangzhou National Laboratory, Guangzhou, China
Haopeng Lin, Guangzhou National Laboratory, Guangzhou, China
Tao Xu, Guangzhou National Laboratory, Guangzhou, China
Lihua Chen, Guangzhou National Laboratory, Guangzhou, China
Lingqiang Zhu, Department of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
Huisheng Liu, Guangzhou National Laboratory, Guangzhou, China
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