Cryopreservation of rat MSCs by use of a programmed freezer with magnetic field

• Published in: Cryobiology Vol. 67; no. 3; pp. 258 - 263
• Main Authors: Kojima, Shunichi, Kaku, Masato, Kawata, Toshitsugu, Sumi, Hiromi, Shikata, Hanaka, Abonti, Tahsin Raquib, Kojima, Shotoku, Fujita, Tadashi, Motokawa, Masahide, Tanne, Kazuo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01-12-2013
• Subjects: Animals; CAS freezer; Cell Differentiation; Cell Proliferation; Cell Survival; Cells, Cultured; Cryopreservation; Cryopreservation - instrumentation; Cryoprotective Agents - chemistry; Dimethyl Sulfoxide - chemistry; Ice - analysis; Magnetic field; Magnetic Fields; Male; Mesenchymal Stromal Cells - cytology; MSCs; Rats; Rats, Inbred F344; CAS freezer; MSCs; Cryopreservation; Magnetic field
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/j.cryobiol.2013.08.003

Mesenchymal stem cells (MSCs) can be used for the regeneration of various tissues and cryopreservation of MSCs is so important for regenerative medicine. The purpose of this study was to evaluate the influences of cryopreservation on MSCs by use of a programmed freezer with a magnetic field (CAS freezer). MSCs were isolated from bone marrow of rat femora. The cells were frozen by a CAS freezer with 10% dimethyl sulfoxide (Me2SO) and cryopreserved for 7days at a temperature of −150°C. Immediately after thawing, the number of survived cells was counted. The cell proliferation also examined after 48h culture. Next, MSCs were frozen by two different freezers; CAS freezer and a conventional programmed freezer without magnetic field. Then, osteogenic and adipogenic differentiations of cryopreserved cells were examined. As a result, survival and proliferation rates of MSCs were significantly higher in CAS freezer than in the non-magnetic freezer. Alizarin positive reaction, large amount of calcium quantification, and greater alkaline phosphatase activity were shown in both the non-cryopreserved and CAS groups after osteogenic differentiation. Moreover, Oil Red O staining positive reaction and high amount of PPARγ and FABP4 mRNAs were shown in both the non-cryopreserved and CAS groups after adipogenic differentiation. From these findings, it is shown that a CAS freezer can maintain high survival and proliferation rates of MSCs and maintain both adipogenic and osteogenic differentiation abilities. It is thus concluded that CAS freezer is available for cryopreservation of MSCs, which can be applied to various tissue regeneration.