title: Establishment of a three-dimensional culture system of gastric stem cells supporting mucous cell differentiation using microfibrous polycaprolactone scaffolds.
abstract: OBJECTIVE
  To generate various polycaprolactone (PCL) scaffolds and test their suitability for growth and differentiation of immortalized mouse gastric stem (mGS) cells.
  
  
  METHODS
  Non-porous, microporous and three-dimensional electrospun microfibrous PCL scaffolds were prepared and characterized for culture of mGS cells. First, growth of mGS cells was compared on these different scaffolds after 3 days culture, using viability assay and microscopy. Secondly, growth pattern of the cells on microfibrous scaffolds was studied after 3, 6, 9 and 12 days culture using DNA PicoGreen assay and scanning electron microscopy. Thirdly, differentiation of the cells grown on microfibrous scaffolds for 3 and 9 days was analysed using lectin/immunohistochemistry.
  
  
  RESULTS
  The mGS cells grew preferentially on microfibrous scaffolds. From 3 to 6 days, there was increase in cell number, followed by reduction by days 9 and 12. To test whether the reduction in cell number was associated with cell differentiation, cryosections of cell-containing scaffolds cultured for 3 and 9 days were probed with gastric epithelial cell differentiation markers. On day 3, none of the markers examined bound to the cells. However by day 9, approximately, 50% of them bound to N-acetyl-d-glucosamine-specific lectin and anti-trefoil factor 2 antibodies, indicating their differentiation into glandular mucus-secreting cells.
  
  
  CONCLUSIONS
  Microfibrous PCL scaffolds supported growth and differentiation of mGS cells into mucus-secreting cells. These data will help lay groundwork for future experiments to explore use of gastric stem cells and PCL scaffolds in stomach tissue engineering.
authors.affiliation: Department of Anatomy, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates; Department of Chemistry, College of Science, United Arab Emirates University, Al Ain, United Arab Emirates.
authors.forename: S
authors.initials: S
authors.lastname: Pulikkot
authors.forename: Y E
authors.initials: YE
authors.lastname: Greish
authors.forename: A-H I
authors.initials: AH
authors.lastname: Mourad
authors.forename: S M
authors.initials: SM
authors.lastname: Karam
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D051379:Mice; D008855:Microscopy, Electron, Scanning; D011091:Polyesters / Q000737:chemistry / Q000494:pharmacology*; D016062:Porosity; D013234:Stem Cells / Q000166:cytology*; D013270:Stomach / Q000166:cytology; D013718:Tensile Strength; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D011091:Polyesters; C016240:polycaprolactone