Active control of the nucleation temperature enhances freezing survival of multipotent mesenchymal stromal cells.
Cryopreservation is a technique that has been extensively used for storage of multipotent mesenchymal stromal cells (MSCs) in regenerative medicine. Therefore, improving current cryopreservation procedures in terms of increasing cell viability and functionality is important. In this study, we optimized the cryopreservation protocol of MSCs derived from the common marmoset Callithrix jacchus (cj), which can be used as a non-human primate model in various pathological and transplantation studies and have a great potential for regenerative medicine. We have investigated the effect of the active control of the nucleation temperature using induced nucleation at a broad range of temperatures and two different dimethylsulfoxide concentrations (Me2SO, 5% (v/v) and 10%, (v/v)) to evaluate the overall effect on the viability, metabolic activity and recovery of cells after thawing. Survival rate and metabolic activity displayed an optimum when ice formation was induced at -10 °C. Cryomicroscopy studies indicated differences in ice crystal morphologies as well as differences in intracellular ice formation with different nucleation temperatures. High subzero nucleation temperatures resulted in larger extracellular ice crystals and cellular dehydration, whereas low subzero nucleation temperatures resulted in smaller ice crystals and intracellular ice formation.
Institute for Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany. Electronic address: lauterboeck@imp.uni-hannover.de.
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Lauterboeck
Institute for Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany. Electronic address: hofmann@imp.uni-hannover.de.
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Hofmann
Service Unit Embryonic Stem Cells, Institute for Transfusion Medicine, Medical School Hannover, Germany. Electronic address: mueller.thomas@mh-hannover.de.
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Mueller
Institute for Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany. Electronic address: glasmacher@imp.uni-hannover.de.
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Glasmacher
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005615:Freezing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D013696:Temperature
D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
Cryomicroscopy; Induced nucleation; Multipotent mesenchymal stromal cells; Reduced Me(2)SO