Importance of a three-stage cooling regime and induced ice nucleation during cryopreservation on colony-forming potential and differentiation in mesenchymal stem progenitor cells from human fetal liver.
Colony-forming activity, as well as osteogenic and adipogenic capacities of primary human fetal liver (HFL) mesenchymal stem or progenitor cells (MSCs) were compared before and after cryopreservation using a standard three-step cooling protocol (Cryo3-S) or the same protocol with induced ice nucleation (Cryo3-IIN) and 5% and 10% w/v dimethyl sulphoxide (Me₂SO). Cell viability, using the Cryo3-S protocol with 5 % and 10 % Me₂SO, was about 60 to 70 % as assessed by the trypan blue staining method, but the ability to undergo growth in culture and form colonies was completely lost. Cryopreservation using Cryo3-IIN resulted in conservation of colony-forming MSCs. Colony-forming efficiency (CFE) of the cell samples cryopreserved with Cryo3-IIN and 5 % Me₂SO was on average 0.4 +/- 0.1 colonies per 10⁵ cells, whereas with 10% Me₂SO 1.6 +/- 0.7 colonies were obtained. HFL MSCs recovered after cryopreservation in the both groups demonstrated capacity to be expanded and induced into either osteogenic or adipogenic differentiation.
Department of Biochemistry of Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
Nataliya  G
N
Skorobogatova
Aalexander N
AN
Novikov
Barry J
BJ
Fuller
Aalexander Y
AY
Petrenko
D002470:Cell Survival; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D015925:Cryopreservation / Q000379:methods*; D053686:Fetal Stem Cells / Q000166:cytology; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000196:embryology; D059630:Mesenchymal Stem Cells / Q000166:cytology*