title: Positive selection of Wharton's jelly-derived CD105(+) cells by MACS technique and their subsequent cultivation under suspension culture condition: A simple, versatile culturing method to enhance the multipotentiality of mesenchymal stem cells.
abstract: OBJECTIVE
  Wharton's jelly (WJ), an appropriate source of mesenchymal stem cells (MSCs), has been shown to have a wide array of therapeutic applications. However, the WJ-derived MSCs are very heterogeneous and have limited expression of pluripotency markers. Hence, improvement of their culture condition would promote the efficiency of WJ-MSCs. This study aims to employ a simple method of cultivation to obtain WJ-MSCs which express more pluripotency markers.
  
  
  METHODS
  CD105(+) cells were separated by magnetic-associated (activated) cell sorting from umbilical cord mucous tissue. CD105(+) cells were added to Methocult medium diluted in α-minimum essential medium (α-MEM) and seeded in poly(2-hydroxyethyl methacrylate) (poly-HEMA)-coated plates for suspension culture preparation. Differentiation capacity of isolated cells was evaluated in the presence of differentiation-inducing media. The expression of pluripotency markers such as Oct3/4, Nanog, and Sox2 was also analyzed by RT-PCR and western blot techniques. Moreover, immunocytochemistry was performed to detect alpha-smooth muscle actin (antigene) (α-SMA) protein.
  
  
  RESULTS
  WJ-MSCs grew homogeneously and formed colonies when cultured under suspension culture conditions (Non-adhesive WJ-MSCs). They maintained their growth ability in both adherent and suspension cultures for several passages. Non-adhesive WJ-MSCs expressed Oct3/4, Nanog, and Sox2 both at transcriptional and translational levels in comparison to those cultured in conventional adherent cultures. They also expressed α-SMA protein.
  
  
  CONCLUSIONS
  In this study, we isolated WJ-MSCs using a slightly modified culture condition. Our simple non-genetic method resulted in a homogeneous population of WJ-MSCs, which highly expressed pluripotency markers.
  
  
  CONCLUSIONS
  In the future, more multipotent WJ-MSCs can be harnessed as a non-embryonic source of MSCs in MSC-based cell therapy.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Amiri
authors.forename: Raheleh
authors.initials: R
authors.lastname: Halabian
authors.forename: Mozhgan
authors.initials: M
authors.lastname: Dehgan Harati
authors.forename: Marzie
authors.initials: M
authors.lastname: Bahadori
authors.forename: Ahmad
authors.initials: A
authors.lastname: Mehdipour
authors.forename: Amaneh
authors.initials: A
authors.lastname: Mohammadi Roushandeh
authors.forename: Mehryar
authors.initials: M
authors.lastname: Habibi Roudkenar
mesh_terms: D015703:Antigens, CD / Q000032:analysis*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002458:Cell Fractionation / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D000071063:Endoglin; D005260:Female; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008689:Methacrylates / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000032:analysis*; D059631:Wharton Jelly / Q000166:cytology*
chemical_list: D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D008689:Methacrylates; D011956:Receptors, Cell Surface; C005044:hydroxyethyl methacrylate
keywords: Cell therapy; MSCs; Pluripotency markers; Wharton's jelly