title: Cryopreservation of rat MSCs by use of a programmed freezer with magnetic field.
abstract: 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 7 days at a temperature of -150 °C. Immediately after thawing, the number of survived cells was counted. The cell proliferation also examined after 48 h 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.
authors: Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University, Graduate School of Biomedical Science, Japan. Electronic address: shunichi0202@hiroshima-u.ac.jp.
authors: Shunichi
authors: S
authors: Kojima
authors: Masato
authors: M
authors: Kaku
authors: Toshitsugu
authors: T
authors: Kawata
authors: Hiromi
authors: H
authors: Sumi
authors: Hanaka
authors: H
authors: Shikata
authors: Tahsin Raquib
authors: TR
authors: Abonti
authors: Shotoku
authors: S
authors: Kojima
authors: Tadashi
authors: T
authors: Fujita
authors: Masahide
authors: M
authors: Motokawa
authors: Kazuo
authors: K
authors: Tanne
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000295:instrumentation*; D003451:Cryoprotective Agents / Q000737:chemistry; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D007053:Ice / Q000032:analysis; D060526:Magnetic Fields; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D011916:Rats, Inbred F344
chemical_list: D003451:Cryoprotective Agents; D007053:Ice; D004121:Dimethyl Sulfoxide
keywords: CAS freezer; Cryopreservation; MSCs; Magnetic field