从人类多能干细胞生成胰腺三角洲细胞的分化方案。

IF 4.6 2区 生物学 Q2 CELL BIOLOGY
Frontiers in Cell and Developmental Biology Pub Date : 2024-10-18 eCollection Date: 2024-01-01 DOI:10.3389/fcell.2024.1490040
Tongran Zhang, Nannan Wang, Zhiying Liao, Jingyi Chen, Hao Meng, Haopeng Lin, Tao Xu, Lihua Chen, Ling-Qiang Zhu, Huisheng Liu
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引用次数: 0

摘要

在本方案中,我们详细介绍了用人多能干细胞(hPSCs)生成胰腺三角洲细胞(SC-三角洲细胞)的七阶段分化方法。第一步,通过活化素A和CHIR99021生成明确的内胚层,然后通过FGF7、SANT1、LDN193189、PdBU和维甲酸(RA)诱导原始肠管和后前肠。在第 4 和第 5 阶段,FGF7 与 FGF2 以及 RA、XXI、ALK5 抑制剂 II、SANT1、Betacellulin 和 LDN193189 的联合处理可实现随后的内分泌生成和定向 SC-delta 细胞诱导。第 5 阶段后,平面培养转换为悬浮系统,使细胞聚集成含有 delta 细胞的球体。产生三角细胞的分化过程大约需要 4-5 周,细胞扩增和评估则需要 1-2 周。我们相信,这种简便易行的方案能为高效分化提供稳定的 SC 三角洲细胞来源,有助于进一步研究三角洲细胞的生理作用,并完善胰岛细胞治疗策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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 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.

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来源期刊
Frontiers in Cell and Developmental Biology
Frontiers in Cell and Developmental Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
9.70
自引率
3.60%
发文量
2531
审稿时长
12 weeks
期刊介绍: Frontiers in Cell and Developmental Biology is a broad-scope, interdisciplinary open-access journal, focusing on the fundamental processes of life, led by Prof Amanda Fisher and supported by a geographically diverse, high-quality editorial board. The journal welcomes submissions on a wide spectrum of cell and developmental biology, covering intracellular and extracellular dynamics, with sections focusing on signaling, adhesion, migration, cell death and survival and membrane trafficking. Additionally, the journal offers sections dedicated to the cutting edge of fundamental and translational research in molecular medicine and stem cell biology. With a collaborative, rigorous and transparent peer-review, the journal produces the highest scientific quality in both fundamental and applied research, and advanced article level metrics measure the real-time impact and influence of each publication.
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