Stepwise differentiation of human embryonic stem cells into early endoderm derivatives and their molecular characterization

Oded Kopper, Nissim Benvenisty*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Human embryonic stem cells have the potential to differentiate into all human cell types and therefore hold a great therapeutic promise. Differentiation into the embryonic endoderm and its derivatives is of special interest since it can provide a cure for severe widespread clinical conditions such as diabetes and hepatic failure. In this work we established a unique experimental outline that enables the study of early human endoderm development and can help improve and create new differentiation protocols. To this end we started with mesendoderm cells and separated them into early endoderm and mesoderm progenitor cells using CXCR4 and PDGFRA cell surface markers. We molecularly characterized the different lineages, and demonstrated the importance of the TGFβ pathway in definitive endoderm initiation. The endoderm progenitor cells were then purified creating an endodermal differentiation niche that is not affected by other cell populations. We followed the differentiation of these cells at different time points, and demonstrated an up regulation of genes indicative to differentiation into both foregut and hindgut. Surprisingly, upon continued culture, there was significant down regulation of the hepatic gene signature. This down regulation could be rescued with FGF2 treatment demonstrating its importance in hepatic cell maintenance. In conclusion, we suggest that isolating endoderm progenitor cells is crucial for the analysis of their fate, and enables the identification of factors involved in their differentiation and maintenance.

Original languageAmerican English
Pages (from-to)335-345
Number of pages11
JournalStem Cell Research
Volume8
Issue number3
DOIs
StatePublished - May 2012

Bibliographical note

Funding Information:
We thank Dr. Danny Kitsberg for kindly providing the primers for the real time SYBR green reactions. We also thank Nadav Sharon and Dr. Yoav Mayshar for critically reading the manuscript. N.B. is the Herbert Cohn Chair in Cancer Research. This research was partially supported by funds from the ISF-Morasha Foundation (grant no. 1801/10 ) to N.B. We gratefully acknowledge support for this project provided by a grant from the Legacy Heritage Fund of New York to N.B.

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