title: A comparative study on culture conditions and routine expansion of amniotic fluid-derived mesenchymal progenitor cells.
abstract: BACKGROUND
  Amniotic fluid (AF) cell populations will be applied in perinatology. We aimed to test the feasibility of large-scale cell expansion.
  
  
  METHODS
  We determined the best out of three published expansion protocols for mesenchymal progenitors (AF samples, n = 4) in terms of self-renewal ability. Characterization was performed based on morphology, surface marker analysis, cytogenetic stability, and differentiation potential. The conditions for the best self-renewal ability were further determined in a consecutive series (n = 159).
  
  
  RESULTS
  The medium containing fetal bovine serum (FBS), epidermal growth factor, insulin, transferrin, and tri-iodothyronine, combined with seeding on gelatin-coated wells, best stimulated the growth of cells with mesenchymal features, as demonstrated by flow cytometry; however, only osteogenic differentiation was possible. Large-scale testing (n = 44) failed to confirm a robust self-renewal ability. Better results were obtained (n = 88) using optimized FBS or an increased initial cell density. Eventually over 81% of cultures continued growing after the initial medium change and had mesenchymal features but failed differentiation assays.
  
  
  CONCLUSIONS
  Routine in vitro expansion of AF-derived mesenchymal cells remains problematic. Despite an increase in successful cell cultures from 40 up to 80% using optimized serum and an increased cell density, eventually cells failed to demonstrate differentiation abilities. Routine isolation and expansion from unselected AF samples remains a challenge.
authors.affiliation: Cluster Organ Systems, University Hospitals Leuven, Leuven, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Gucciardo
authors.forename: N
authors.initials: N
authors.lastname: Ochsenbein-Kölble
authors.forename: Y
authors.initials: Y
authors.lastname: Ozog
authors.forename: G
authors.initials: G
authors.lastname: Verbist
authors.forename: V
authors.initials: V
authors.lastname: Van Duppen
authors.forename: J P
authors.initials: JP
authors.lastname: Fryns
authors.forename: R
authors.initials: R
authors.lastname: Lories
authors.forename: J
authors.initials: J
authors.lastname: Deprest
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002417:Cattle; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D003470:Culture Media; D005260:Female; D006801:Humans; D007621:Karyotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011247:Pregnancy
chemical_list: D003470:Culture Media