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KMID : 0894520110150040281
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Development & Reproduction
2011 Volume.15 No. 4 p.281 ~ p.289
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Study of Effective Cryoprotectants on the Cryopreservation of Porcine Mesenechymal Stem Cells
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Kim Mi-Kyeong
Park Hyoung-Joon
Rho Gyu-Jin
Cho Jae-Hyeon
Kim Chung-Hui
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Abstract
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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°C/min from 25°C up to -80°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.
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KEYWORD
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Porcine mesenchymal stem cells, Cryopreservation, Dimethyl sulfoxide, Ethylene Glycol, Cell viability, Gene expression
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