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首页> 外文期刊>Research Opinions in Animal & Veterinary Sciences >Assessment of the effect of different concentration of foetal bovine serum (FBS) in freezing media on the viability of spermatogonial stem cells
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Assessment of the effect of different concentration of foetal bovine serum (FBS) in freezing media on the viability of spermatogonial stem cells

机译：评估冷冻培养基中不同浓度的胎牛血清（FBS）对精原干细胞活力的影响

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An in vitro system that supports spermatogonial stem cells survival and proliferation is useful for enhancement of stem cell number and efficient transplantation. The purpose of this study was to assess the effect of different concentrations of foetal bovine serum on survival of bovine spermatogonial stem cells and Sertoli cells after cryo- preservation. Samples were surgically obtained from testes of 3-5 months old calves and digested twice in enzymatic media to get the isolated cells. These cells were co- cultured for two weeks to obtain the adequate amount of cells. In the freezing step, the purified spermatogonial stem cells were frozen in four different freezing media including bio-freeze medium . as Control and 3 other groups containing: Dulbecco's modified Eagle's medium (DMEM) and 50%, 70% and 90% fetal bovine serum (FBS) plus 10% dimethyl-sulfoxide (DMSO) as treatments. After 2 weeks of preserving in liquid nitrogen, to assess the viability, the cells were rapidly thawed by swirling in 38°C water bath for two minutes. The viability of the cells was assessed by using flow cyto metry technique. The results showed that the frozen cells in 50 and 70% FBS groups had significantly (P<0.05) higher viability rates than control and 90% FBS groups. It is concluded that spermatogonial stem cells can be cryopreserved in DMEM plus 10% DMSO and 50 to 70% FBS.
机译：支持精原干细胞存活和增殖的体外系统对于增加干细胞数量和有效移植是有用的。这项研究的目的是评估冷冻保存后不同浓度的胎牛血清对牛精原干细胞和支持细胞存活的影响。从3-5个月大的小牛睾丸中手术获取样品，并在酶促培养基中消化两次以得到分离的细胞。将这些细胞共培养两周以获得足够量的细胞。在冷冻步骤中，将纯化的精原干细胞在四种不同的冷冻培养基中冷冻，包括生物冷冻培养基。作为对照组，另3个组包括：Dulbecco改良的Eagle培养基（DMEM）和50％，70％和90％的胎牛血清（FBS）加10％的二甲基亚砜（DMSO）作为治疗。在液氮中保存2周后，为了评估其生存能力，将细胞在38°C水浴中涡旋2分钟，迅速融化。通过使用流式细胞仪技术评估细胞的生存力。结果表明，与对照组和90％FBS组相比，在50％和70％FBS组中，冷冻细胞的存活率显着提高（P <0.05）。结论是可以将精原干细胞冷冻保存在DMEM加10％DMSO和50％至70％FBS中。

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