Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (7): 1439-1446.doi: 10.3864/j.issn.0578-1752.2012.07.024

• RESEARCH NOTES • Previous Articles     Next Articles

Effects of Different Cryoprotectants on Ovine Preadipocytes Cryopreservation

 CAI  Yong, A  Yi-Mu-Gu-Li, ZANG  Rong-Xin, LIU  Yi-Zhong, YANG  Ju-Tian, QIAO  Zi-Lin, CAO  Xin, XU  Hong-Wei, WU  Jian-Ping   

  1. 1.西北民族大学实验中心，兰州 730030
    2.甘肃农业大学动物科学技术学院/国家绒毛用羊产业体系环境控制实验室，兰州 730070
    3.西北民族大学生命科学与工程学院，兰州 730030
    ４.甘肃省动物细胞工程技术研究中心，兰州 730030
  • Received:2011-03-09 Online:2012-04-01 Published:2011-10-11

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Abstract: 【Objective】 The aim of this experiment is to investigate a suitable cryoprotectant and its optimal concentration for ovine preadipocytes from omental. 【Method】Trypan blue exclusion test, MTT, oil red O staining and real-time PCR were used to test the effects of following cryoprotective agents (CPAs) with different concentrations on post-thaw survival, proliferation, differentiation capacity, PPARγ and LPL mRNA expression of ovine preadipocytes from omental, dimethyl sulphoxide (DMSO), ethylene glycol (EG), propylene glycol (PG), glycerol (G) and polyvinylpyrrolidone (PVP). In addition to the CPAs, the basic medium is DMEM/F12 medium plus 20% FBS. Then karyotype was analyzed. 【Result】Trypan blue exclusion test showed that the highest survival rate, no significant difference with the primary cells, was obtained when cryopreserved with 10% DMSO or 5% DMSO plus 5% PVP among all the CPA treatments in this study. The highest survival rate (94.96%) and cell viability were obtained when cryopreserved with 10% PVP, and showed no significant difference compared with primary cells. Oil red O staining showed no significant difference in lipogenesis among all the CPAs groups and primary cells on 6 th day (P＞0.05), while on 11th day, cells cryopreserved with 10% DMSO turned up significantly greater lipogenesis than other CPAs (P＜0.05), but had no significant difference with primary cells (P＞0.05). Activity of the GPDH, mRNA expression of LPL and PPARγ showed no significant difference among all the groups (P＞0.05). Karyotype analysis showed that diploid cells were dominant post thaw and showed no significant difference compared with primary cells (P＞0.05). 【Conclusion】Results of the present study indicate that ovine preadipocytes could successfully be cryopreserved with DMEM/F12 medium containing 10% DMSO or 10 % PVP, 5% DMSO plus 5% PVP, while 10% DMSO is a suitable CPA for ovine preadipocytes.

Key words: cryopreservation, preadipocyte, ovine, cryoprotectants

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