Open AccessPurification of Live Stem‐Cell‐Derived Islet Lineage Intermediates
Author(s) -
Rasouli Niloofar,
Melton Douglas A.,
AlvarezDominguez Juan R.
Publication year - 2020
Publication title -
current protocols in stem cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.658
H-Index - 28
eISSN - 1938-8969
pISSN - 1941-7322
DOI - 10.1002/cpsc.111
Subject(s) - biology , stem cell , progenitor cell , enteroendocrine cell , islet , flow cytometry , microbiology and biotechnology , lineage (genetic) , endocrine system , in vitro , insulin , endocrinology , gene , hormone , biochemistry
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 + β 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