• Citer

Safe and effective cryopreservation methods for long-term storage of human-amniotic-fluid-derived stem cells

par Hennes, Aurélie;Deprest, Jan A M J.A.;Toelen, Jaan;Gucciardo, Leonardo ;Zia, Silvia;Lesage, Flore;Lefevre, Nicolas ;Lewi, Liesbeth;Vorsselmans, Anniek;Cos, Theresa;Lories, Rik
Référence Prenatal diagnosis, 35, 5, page (456-462)
Publication Publié, 2015-05

Article révisé par les pairs

• Accès en ligne
• Détails
• Contenu
• Statistiques

Résumé :

What's already known about this topic? Stem cells can be isolated from amniotic fluid and may be used for cell therapy and tissue engineering applications in perinatal medicine. Clinical implementation requires development of an adapted cell banking system including optimal processing and cryopreservation of amniotic fluid samples, with maintenance of cell viability and characteristics after thawing. What does this study add? We identified three suitable cryopreservation protocols of fresh amniotic fluid samples with high cell recovery rate and unchanged stem cell characteristics post-thawing. One of these is compatible with current good manufacturing practice legislation and can be performed in fetal medicine units without stem cell culture expertise. Objectives: 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.