title: Defined three-dimensional culture conditions mediate efficient induction of definitive endoderm lineage from human umbilical cord Wharton's jelly mesenchymal stem cells.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Research Division, Dasman Diabetes Institute, 1180, Dasman, Kuwait. ashraf.madhoun@dasmaninstitute.org.
authors.forename: Ashraf
authors.identifier: 0000-0001-8593-3878
authors.initials: A
authors.lastname: Al Madhoun
authors.affiliation: Research Division, Dasman Diabetes Institute, 1180, Dasman, Kuwait.|Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Health Sciences Center, Kuwait University, Al-Jabriya, Kuwait.
authors.forename: Hamad
authors.initials: H
authors.lastname: Ali
authors.affiliation: Research Division, Dasman Diabetes Institute, 1180, Dasman, Kuwait.
authors.forename: Sarah
authors.initials: S
authors.lastname: AlKandari
authors.affiliation: Research Division, Dasman Diabetes Institute, 1180, Dasman, Kuwait.
authors.forename: Valerie Lopez
authors.initials: VL
authors.lastname: Atizado
authors.affiliation: Research Division, Dasman Diabetes Institute, 1180, Dasman, Kuwait.
authors.forename: Nadeem
authors.initials: N
authors.lastname: Akhter
authors.affiliation: Department of Pathology, Molecular Pathology Unit, Faculty of Medicine, Health Sciences Center, Kuwait University, Al-Jabriya, Kuwait.
authors.forename: Fahd
authors.initials: F
authors.lastname: Al-Mulla
authors.affiliation: UIC Regenerative Medicine Research Institute, International University of Catalonia, Barcelona, Spain.
authors.forename: Maher
authors.initials: M
authors.lastname: Atari
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000378:metabolism; D004707:Endoderm / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D014470:Umbilical Cord / Q000166:cytology* / Q000378:metabolism; D059631:Wharton Jelly / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D016895:Culture Media, Serum-Free; D036341:Intercellular Signaling Peptides and Proteins
keywords: 3D; CXCR4; Definitive endoderm; Mesenchymal stromal cells; Sox17; Wharton’s jelly; Wharton’s jelly-derived mesenchymal stem cells