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Study of Effective Cryoprotectants on the Cryopreservation of Porcine Mesenechymal Stem Cells  

Kim, Mi-Kyeong (Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University)
Park, Hyoung-Joon (Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University)
Rho, Gyu-Jin (Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University)
Kim, Chung-Hei (Dept. of Animal Science & Biotechnology, Gyeongnam National University of Science and Technology)
Cho, Jae-Hyeon (Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University)
Publication Information
Development and Reproduction / v.15, no.4, 2011 , pp. 281-289 More about this Journal
Abstract
The objective of this study was to investigate the effective cryoprotectants for the cryopreservation of porcine mesenechymal stem cells (pMSCs). In order to understand the effectiveness of various cryoprotectants on pMSCs, we studied the most commonly used cryoprotectants; dimethyl sulfoxide (DMSO), ethylene glycol (EG), DMSO and EG. pMSCs were isolated from bone marrow matrix of piglet (2 month) and characterized by alkaline phopshatase (AP) activity, colony forming, and differentiation to adipocyte. In slow cooling cryopreservation, the pMSCs were exposed to cell medium containing Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% DMSO, 1.5M EG and 5% DMSO/0.75M EG, respectively, and freezed to $-1^{\circ}C$/min from $25^{\circ}C$ up to $-80^{\circ}C$ in a cryo-container. The proportion of viable cells and the growing rates in fresh pMSCs were significantly (P<0.05) higher than those of other groups, but did not differ between the cryopreserved groups. The expression of Sox-2 and Nanog gene was increased by extending culture time in cryopreserved groups. The expression of Bax gene in cryopreserved groups was similar with fresh pMSCs. Moreover, the gene expression of adipocyte-specific marker as well as chondrogenic/osteogenic factors in cryopreserved groups was similarly to fresh pMSCs. Taken together, our results suggested that all these cryoprotectants of 10% DMSO, 1.5M EG and 5% DMSO/0.75M EG could be used for cryopreservation of the pMSCs.
Keywords
Porcine mesenchymal stem cells; Cryopreservation; Dimethyl sulfoxide; Ethylene Glycol; Cell viability; Gene expression;
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