A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells
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 post...
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Frontiers Media S.A.
2024-10-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1490040/full |
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| author | Tongran Zhang Tongran Zhang Nannan Wang Nannan Wang Zhiying Liao Zhiying Liao Jingyi Chen Jingyi Chen Hao Meng Hao Meng Haopeng Lin Tao Xu Tao Xu Lihua Chen Lihua Chen Ling-Qiang Zhu Huisheng Liu Huisheng Liu Huisheng Liu Huisheng Liu |
| author_facet | Tongran Zhang Tongran Zhang Nannan Wang Nannan Wang Zhiying Liao Zhiying Liao Jingyi Chen Jingyi Chen Hao Meng Hao Meng Haopeng Lin Tao Xu Tao Xu Lihua Chen Lihua Chen Ling-Qiang Zhu Huisheng Liu Huisheng Liu Huisheng Liu Huisheng Liu |
| author_sort | Tongran Zhang |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-cad4d229a56d4bdeb54fb65e41d30b16 |
| institution | OA Journals |
| issn | 2296-634X |
| language | English |
| publishDate | 2024-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj-art-cad4d229a56d4bdeb54fb65e41d30b162025-08-20T01:47:50ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2024-10-011210.3389/fcell.2024.14900401490040A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cellsTongran Zhang0Tongran Zhang1Nannan Wang2Nannan Wang3Zhiying Liao4Zhiying Liao5Jingyi Chen6Jingyi Chen7Hao Meng8Hao Meng9Haopeng Lin10Tao Xu11Tao Xu12Lihua Chen13Lihua Chen14Ling-Qiang Zhu15Huisheng Liu16Huisheng Liu17Huisheng Liu18Huisheng Liu19Department of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaCollege of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaSchool of Biomedical Engineering, Guangzhou Medical University, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaSchool of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaSchool of Biomedical Engineering, Guangzhou Medical University, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaSchool of Biomedical Engineering, Guangzhou Medical University, Guangzhou, Guangdong, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaSchool of Biomedical Engineering, Guangzhou Medical University, Guangzhou, Guangdong, ChinaDepartment of Pathophysiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaDepartment of Testing and Diagnosis Technology Research, Guangzhou National Laboratory, Guangzhou, Guangdong, ChinaCollege of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaSchool of Biomedical Engineering, Guangzhou Medical University, Guangzhou, Guangdong, ChinaSchool of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou, Guangdong, ChinaIn 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.https://www.frontiersin.org/articles/10.3389/fcell.2024.1490040/fullhPSCsstem cell differentiationpancreatic delta cellsFGFislet organoids |
| spellingShingle | Tongran Zhang Tongran Zhang Nannan Wang Nannan Wang Zhiying Liao Zhiying Liao Jingyi Chen Jingyi Chen Hao Meng Hao Meng Haopeng Lin Tao Xu Tao Xu Lihua Chen Lihua Chen Ling-Qiang Zhu Huisheng Liu Huisheng Liu Huisheng Liu Huisheng Liu A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells Frontiers in Cell and Developmental Biology hPSCs stem cell differentiation pancreatic delta cells FGF islet organoids |
| title | A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| title_full | A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| title_fullStr | A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| title_full_unstemmed | A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| title_short | A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| title_sort | differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells |
| topic | hPSCs stem cell differentiation pancreatic delta cells FGF islet organoids |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1490040/full |
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