Defined three-dimensional culture conditions mediate efficient induction of definitive endoderm lineage from human umbilical cord Wharton's jelly mesenchymal stem cells

Ashraf Al Madhoun, Hamad Mohamed Ali Yaseen, Sarah Alkandari, Valerie Lopez Atizado, Nadeem Akhter, Fahd Al-Mulla, Maher Atari

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Background: Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are gaining increasing interest as an alternative source of stem cells for regenerative medicine applications. Definitive endoderm (DE) specification is a prerequisite for the development of vital organs such as liver and pancreas. Hence, efficient induction of the DE lineage from stem cells is crucial for subsequent generation of clinically relevant cell types. Here we present a defined 3D differentiation protocol of WJ-MSCs into DE cells. Methods: WJ-MSCs were cultured in suspension to generate spheroids, about 1500 cells each, for 7 days. The serum-free differentiation media contained specific growth factors, cytokines, and small molecules that specifically regulate signaling pathways including sonic hedgehog, bone morphogenetic protein, Activin/Wnt, and Notch. Results: We obtained more than 85 % DE cells as shown with FACS analysis using antibodies directed against the DE marker CXCR4. In addition, biochemical and molecular analysis of bona-fide DE markers revealed a time-course induction of Sox17, CXCR4, and FoxA2. Focused PCR-based array also indicated a specific induction into the DE lineage. Conclusions: In this study, we report an efficient serum-free protocol to differentiate WJ-MSCs into DE cells utilizing 3D spheroid formation. Our approach might aid in the development of new protocols to obtain DE-derivative lineages including liver-like and pancreatic insulin-producing cells.

Original languageEnglish
Article number165
JournalStem Cell Research and Therapy
Volume7
Issue number1
DOIs
Publication statusPublished - 16 Nov 2016

Keywords

  • 3D
  • CXCR4
  • Definitive endoderm
  • Mesenchymal stromal cells
  • Sox17
  • Wharton's jelly
  • Wharton's jelly-derived mesenchymal stem cells

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