Efficient Cryopreservation of Human Pluripotent Stem Cells by Surface-Based Vitrification
Efficient cryopreservation of human stem cells is crucial for guaranteeing a permanent supply of high-quality cell material for drug discovery or regenerative medicine. Conventionally used protocols usually employing slow freezing rates, however, result in low recovery rates for human pluripotent stem cells due to their complex colony structure. In this chapter, a surface-based vitrification protocol for pluripotent stem cells is presented based on a procedure for human embryonic stem cells developed by Beier et al. (Cryobiology 63:175–185, 2011). This simple and highly efficient cryopreservation method allows cryopreservation of large numbers of ready-to-use adherent cells that maintain pluripotency.
Key wordsCryopreservation Pluripotent stem cells Surface-based vitrification Vitrification
The work with human embryonic stem cells was permitted by the Robert Koch Institute (18 and 44 permission) and carried out according to German law.
- 5.Strulovici Y, Leopold PL, O’Connor TP, Pergolizzi RG, Crystal RG (2007) Human embryonic stem cells and gene therapy. Mol Ther 15:850–866Google Scholar
- 7.Katsen-Globa A, Meiser I, Petrenko YA, Ivanov RV, Lozinsky VI, Zimmermann H, Petrenko AY (2014) Towards ready-to-use 3-D scaffolds for regenerative medicine: adhesion-based cryopreservation of human mesenchymal stem cells attached and spread within alginate-gelatin cryogel scaffolds. J Mater Sci Mater Med 25:857–871CrossRefGoogle Scholar
- 9.Heng BC, Ye CP, Liu H, Toh WS, Rufaihah AJ, Yang Z et al (2006) Loss of viability during freeze-thaw of intact and adherent human embryonic stem cells with conventional slow-cooling protocols is predominantly due to apoptosis rather than cellular necrosis. J Biomed Sci 13:433–445CrossRefGoogle Scholar