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Impact of co‐culture on pancreatic differentiation of embryonic stem cells
Author(s) -
Banerjee Ipsita,
Sharma Nripen,
Yarmush Martin
Publication year - 2011
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.317
Subject(s) - microbiology and biotechnology , embryonic stem cell , downregulation and upregulation , pdx1 , matrigel , cellular differentiation , stem cell , biology , population , islet , embryoid body , endocrinology , chemistry , medicine , insulin , cell , adult stem cell , biochemistry , environmental health , gene
Promise of cellular therapy for type 1 diabetes has inspired the search for transplantable cell sources, and embryonic stem cells (ESCs) have emerged as strong candidates. We have developed a directed differentiation protocol to obtain insulin‐producing cells from ESCs. The ESCs are first induced towards a homogeneous monolayer of definitive endoderm‐like cells by co‐culture with primary hepatocytes. Pancreatic commitment is induced by plating the ESC‐derived endoderms on Matrigel, along with Sonic hedgehog inhibition and retinoid induction. More than 70% of differentiated cells positively upregulated Pdx‐1, along with pro‐endocrine transcription factors Ngn3, β2/neroD1, Nkx2.2 and Nkx6.1. Final maturation to islet‐specific cells is achieved by co‐culturing the ESC‐derived pancreatic endocrine cells with endothelial cells, which resulted in Insulin 1 upregulation in 60% of the cell population, along with high levels of IAPP and Glut2. The differentiated cell population also secreted high levels of insulin. Our findings illustrate the significant effect of co‐culture in different stages of differentiation and maturation of ESCs in vitro . Such a high yield of pancreatic islet cells has not yet been reported. Our findings establish a robust protocol for islet differentiation. Copyright © 2010 John Wiley & Sons, Ltd.