Purification of Live Stem-Cell-Derived Islet Lineage Intermediates.

Current Protocols in Stem Cell Biology
Niloofar RasouliJuan R Alvarez-Dominguez

Abstract

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.

References

May 30, 2013·Development·Felicia W Pagliuca, Douglas A Melton
Aug 6, 2013·Annual Review of Cell and Developmental Biology·Hung Ping ShihMaike Sander
Sep 12, 2014·Nature Biotechnology·Alireza RezaniaTimothy J Kieffer
Oct 11, 2014·Cell·Felicia W PagliucaDouglas A Melton
Aug 25, 2017·Nature Communications·Kathryn F CoggerM Cristina Nostro
May 10, 2019·Nature·Adrian VeresDouglas A Melton
Nov 23, 2019·Current Protocols in Stem Cell Biology·Jeffrey C DavisDouglas A Melton
Dec 17, 2019·Cell Stem Cell·Juan R Alvarez-DominguezDouglas A Melton

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