title: The effects of culture conditions on the functionality of efficiently obtained mesenchymal stromal cells from human cord blood.
abstract: OBJECTIVE
  Cord blood (CB) is an attractive source of mesenchymal stromal cells (MSCs) because of its abundant availability and ease of collection. However, the success rate of generating CB-MSCs is low. In this study, our aim was to demonstrate the efficiency of our previously described method to obtain MSCs from CB and further characterize them and to study the effects of different culture conditions on MSCs.
  
  
  METHODS
  CB-MSC cultures were established in low oxygen (3%) conditions on fibronectin in 10% fetal bovine serum containing culture medium supplemented with combinations of growth factors. Cells were characterized for their adipogenic, osteogenic and chondrogenic differentiation capacity; phenotype; and HOX gene expression profile. The functionality of the cells cultured in different media was tested in vitro with angiogenesis and T-cell proliferation assays.
  
  
  RESULTS
  We demonstrate 87% efficacy in generating MSCs from CB. The established cells had typical MSC characteristics with reduced adipogenic differentiation potential and a unique HOX gene fingerprint. Growth factor-rich medium and a 3% oxygen condition enhanced cell proliferation; however, the growth factor-rich medium had a negative effect on the expression of CD90. Dexamethasone-containing medium improved the capacity of the cells to suppress T-cell proliferation, whereas the cells grown without dexamethasone were more able to support angiogenesis.
  
  
  CONCLUSIONS
  Our results demonstrate that the composition of expansion medium is critical for the functionality of MSCs and should always be appropriately defined for each purpose.
authors: Cell Therapy Services, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
authors: Anita
authors: A
authors: Laitinen
authors: Faculty of Medicine, Department of Pharmacology, University of Helsinki, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
authors: Milla
authors: M
authors: Lampinen
authors: Institute of Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf, Germany.
authors: Stefanie
authors: S
authors: Liedtke
authors: Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
authors: Lotta
authors: L
authors: Kilpinen
authors: Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
authors: Erja
authors: E
authors: Kerkelä
authors: FICAM Finnish Center for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland; Cell Biology, School of Medicine, University of Tampere, Tampere, Finland.
authors: Jertta-Riina
authors: JR
authors: Sarkanen
authors: FICAM Finnish Center for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.
authors: Tuula
authors: T
authors: Heinonen
authors: Institute of Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf, Germany.
authors: Gesine
authors: G
authors: Kogler
authors: Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland. Electronic address: saara.laitinen@bloodservice.fi.
authors: Saara
authors: S
authors: Laitinen
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism / Q000494:pharmacology*; D005312:Fetal Blood / Q000166:cytology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects
chemical_list: D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins
keywords: CD90; angiogenesis; cord blood; immunosuppression; low oxygen; mesenchymal stromal cell