Generation of enhanced definitive endoderm from human embryonic stem cells under an albumin/insulin-free and chemically defined condition

Su Qu, Liang Yan, Bo Fang, Shoudong Ye, Ping Li, Shengyang Ge, Jian Wu, Di Qu, Houyan Song

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Aim To enhance survival and generation of definitive endoderm cells from human embryonic stem cells in a simple and reproducible system. Main methods Definitive endoderm (DE) differentiation from human embryonic stem cells (hESCs) was induced under a chemical-defined condition withdrawn insulin supplement and serum albumin. We dissected influence of “alternative growth factors”, WNT3A, BMP4 and bFGF in activin A-driven differentiation by detection of DE-associated genes expression and cell viability. Expression of DE-associated SOX17 and FOXA2 genes was analyzed by real time reverse transcription polymerase chain reaction (RT-PCR) and Western blot assays. Quantitative evaluation of DE efficiency was performed by flow cytometry analysis of CXCR4-expressed cell population. Cell viability during DE differentiation was analyzed by an Annexin V/PI double staining test. Key findings Supplementation with WNT3A, BMP4 or bFGF promoted DE generation in a dose- and time-dependent manner. Cell apoptosis elicited by activin A was significantly ameliorated by a cocktail with WNT3A, BMP4 and bFGF. This allowed for sustained cell viability without insulin-containing supplements, thereby indirectly improving the efficiency of DE generation. Therefore, the cocktail containing is optimal for efficient DE generation in the presence of activin A and an insulin/albumin-free condition. Significance This optimal condition facilitates the balance between the productivity and the viability maintenance, and could be valuable for mass production of DE with minimal variation.

Original languageEnglish (US)
Pages (from-to)37-46
Number of pages10
JournalLife Sciences
StatePublished - Apr 15 2017
Externally publishedYes


  • Cell survival
  • Chemically defined condition
  • Definitive endoderm
  • Human embryonic stem cell

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)


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