Purification of Live Stem-Cell-Derived Islet Lineage Intermediates.
Purification of Live Stem-Cell-Derived Islet Lineage Intermediates.
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DOI：
10.1002/cpsc.111
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发表时间：
2020-06-01
影响因子：
--
通讯作者：
Alvarez-Dominguez, Juan R
中科院分区：
文献类型：
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作者：
Rasouli, Niloofar;Melton, Douglas A;Alvarez-Dominguez, Juan R
干细胞衍生的组织提供了研究器官发育的平台，健康和疾病期间的生理模型，以及测试新疗法。我们描述了在人干细胞体外分化为胰腺内分泌谱系的过程中在连续阶段分离细胞的方法。使用流式细胞术，我们纯化了胎儿活检无法获得的数量的活谱系中间体。这些包括胰腺和内分泌祖细胞，基于已知的表面标志物分离。我们进一步报告了一种利用细胞内锌含量和DPP 4/CD 26表达将单激素胰岛素+ β细胞与多激素对应物分离的策略。这些方法使得能够在人胰岛谱系进展期间进行全面的分子谱分析。© 2020 Wiley Periodicals LLC.基本方案：人胰岛发育中间体的体外分离。
Stem-cell-derived tissues offer platforms to study organ development, model physiology during health and disease, and test novel therapies. We describe methods to isolate cells at successive stages during in vitro differentiation of human stem cells into the pancreatic endocrine lineage. Using flow cytometry, we purify live lineage intermediates in numbers not available by fetal biopsy. These include pancreatic and endocrine progenitors, isolated based on known surface markers. We further report a strategy that leverages intracellular zinc content and DPP4/CD26 expression to separate monohormonal insulin+ beta cells from polyhormonal counterparts. These methods enable comprehensive molecular profiling during human islet lineage progression. © 2020 Wiley Periodicals LLC. Basic Protocol: In vitro isolation of human islet developmental intermediates.