title: Good manufacturing practice-compliant isolation and culture of human umbilical cord blood-derived mesenchymal stem cells.
abstract: BACKGROUND
  Mesenchymal stem cells (MSCs) are an attractive source of stem cells for clinical applications. These cells exhibit a multilineage differentiation potential and strong capacity for immune modulation. Thus, MSCs are widely used in cell therapy, tissue engineering, and immunotherapy. Because of important advantages, umbilical cord blood-derived MSCs (UCB-MSCs) have attracted interest for some time. However, the applications of UCB-MSCs are limited by the small number of recoverable UCB-MSCs and fetal bovine serum (FBS)-dependent expansion methods. Hence, this study aimed to establish a xenogenic and allogeneic supplement-free expansion protocol.
  
  
  METHODS
  UCB was collected to prepare activated platelet-rich plasma (aPRP) and mononuclear cells (MNCs). aPRP was applied as a supplement in Iscove modified Dulbecco medium (IMDM) together with antibiotics. MNCs were cultured in complete IMDM with four concentrations of aPRP (2, 5, 7, or 10%) or 10% FBS as the control. The efficiency of the protocols was evaluated in terms of the number of adherent cells and their expansion, the percentage of successfully isolated cells in the primary culture, surface marker expression, and in vitro differentiation potential following expansion.
  
  
  RESULTS
  The results showed that primary cultures with complete medium containing 10% aPRP exhibited the highest success, whereas expansion in complete medium containing 5% aPRP was suitable. UCB-MSCs isolated using this protocol maintained their immunophenotypes, multilineage differentiation potential, and did not form tumors when injected at a high dose into athymic nude mice.
  
  
  CONCLUSIONS
  This technique provides a method to obtain UCB-MSCs compliant with good manufacturing practices for clinical application.
authors.affiliation: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh city, Vietnam. pvphuc@hcmuns.edu.vn.
authors.forename: Phuc Van
authors.initials: PV
authors.lastname: Pham
authors.forename: Ngoc Bich
authors.initials: NB
authors.lastname: Vu
authors.forename: Vuong Minh
authors.initials: VM
authors.lastname: Pham
authors.forename: Nhung Hai
authors.initials: NH
authors.lastname: Truong
authors.forename: Truc Le-Buu
authors.initials: TL
authors.lastname: Pham
authors.forename: Loan Thi-Tung
authors.initials: LT
authors.lastname: Dang
authors.forename: Tam Thanh
authors.initials: TT
authors.lastname: Nguyen
authors.forename: Anh Nguyen-Tu
authors.initials: AN
authors.lastname: Bui
authors.forename: Ngoc Kim
authors.initials: NK
authors.lastname: Phan
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects; D000818:Animals; D063646:Carcinogenesis / Q000187:drug effects / Q000473:pathology; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods* / Q000592:standards*; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology*; D019983:Guideline Adherence / Q000592:standards*; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008819:Mice, Nude; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D013997:Time Factors
chemical_list: D003470:Culture Media