Protocol

Pluripotent Stem Cells

Volume 997 of the series Methods in Molecular Biology pp 127-140

Date:

Directed Differentiation of Human Pluripotent Stem Cells Along the Pancreatic Endocrine Lineage

  • Dennis Van HoofAffiliated withDiabetes Center, University of California San Francisco
  • , Muluye E. LikuAffiliated withDiabetes Center, University of California San Francisco

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Abstract

Many research groups are engaged in using human pluripotent stem cells (hPSCs) to generate surrogate pancreatic β-cells for transplantation into diabetic patients. However, to our knowledge, there is no report on the successful generation of glucose-responsive insulin-producing β-cells from hPSCs in vitro. Below, we outline a method that is based on published protocols as well as our own experience by which one can differentiate hPSCs along the pancreatic lineage to generate insulin-producing β-cell-like cells. The protocol, which spans five distinct stages, is an attempt to recapitulate the derivation of pancreatic β-cells in vitro as they form in the developing embryo. We included details on materials and techniques, suggest ways to customize it to your hPSC line of choice, added notes on how to monitor and analyze the cells during differentiation, and indicate what results can be expected.

Key words

Human pluripotent stem cells Definitive endoderm Islets of Langerhans Pancreatic β-cells Insulin-producing cells In vitro differentiation Growth factors Pancreas development