title: Safe and effective cryopreservation methods for long-term storage of human-amniotic-fluid-derived stem cells.
abstract: OBJECTIVE
  Stem cells (SCs) can be isolated from amniotic fluid (AF) for a variety of perinatal applications. In view of this, we compared different cryopreservation protocols for these AFSCs.
  
  
  METHODS
  We screened seven freezing and thawing protocols using two well-established human AFSC lines: freezing protocol 1 (FP1), 10% dimethyl sulfoxide (DMSO); FP2, 2.5% DMSO, caspase inhibitor, and catalase; FP3, 5% glycerol, caspase inhibitor, and catalase; FP4, sperm freezing medium; FP5, slow-freezing solution; FP6, ethylene glycol, sucrose, and Ficoll 70; and FP7, vitrification solution. Outcome measures were post-thawing cell viability, recovery, doubling time and mesenchymal SC markers. The three best performing protocols were subsequently tested on cells isolated from clinical consecutive freshly harvested AF samples from two fetal medicine units.
  
  
  RESULTS
  Protocols 1, 5, and 6 performed significantly better on well-characterized cell lines. They performed equally well on cell pellets from freshly harvested AF (n = 28).
  
  
  CONCLUSIONS
  We identified three suitable cryopreservation protocols because of high cell recovery and unchanged SC characteristics. Given one of these, the slow-freezing solution, is compatible with current good manufacturing practice legislation, it may be ultimately clinically used.
authors: Department of Development and Regeneration, Organ System Cluster, Group Biomedical Sciences, KU, Leuven, Belgium.
authors: Aurélie
authors: A
authors: Hennes
authors: Léonardo
authors: L
authors: Gucciardo
authors: Silvia
authors: S
authors: Zia
authors: Flore
authors: F
authors: Lesage
authors: Nicolas
authors: N
authors: Lefèvre
authors: Liesbeth
authors: L
authors: Lewi
authors: Annick
authors: A
authors: Vorsselmans
authors: Teresa
authors: T
authors: Cos
authors: Rik
authors: R
authors: Lories
authors: Jan
authors: J
authors: Deprest
authors: Jaan
authors: J
authors: Toelen
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D015703:Antigens, CD / Q000378:metabolism; D061945:Caspase Inhibitors; D002374:Catalase; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D004121:Dimethyl Sulfoxide; D019855:Ethylene Glycol; D005260:Female; D005362:Ficoll; D005990:Glycerol; D006801:Humans; D011247:Pregnancy; D012996:Solutions; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D013395:Sucrose; D058989:Vitrification
chemical_list: D015703:Antigens, CD; D061945:Caspase Inhibitors; D012996:Solutions; D005362:Ficoll; D013395:Sucrose; D002374:Catalase; D019855:Ethylene Glycol; D005990:Glycerol; D004121:Dimethyl Sulfoxide