Wang Peng, Li Ying, Hu Xiao-Chen, Cai Xiao-Li, Hou Li-Peng, Wang Yan-Feng, Hu Jian-Hong, Li Qing-Wang, Suo Li-Juan, Fan Zhi-Guo, Zhang Bo
College of Animal Science and Technology, Northwest A & F University, Yangling, Shaanxi 712100, P.R. China.
Zygote. 2014 May;22(2):158-63. doi: 10.1017/S0967199412000378. Epub 2012 Sep 14.
Spermatogonial stem cells (SSCs) have the ability to self-renew and offer a pathway for genetic engineering of the male germ line. Cryopreservation of SSCs has potential value for the treatment of male infertility, spermatogonial transplantation, and so on. In order to investigate the cryopreservation effects of different cryoprotectants on murine SSCs, 0.2 M of low-density lipoproteins (LDL), trehalose and soybean lecithin were added to the cryoprotective medium, respectively, and the murine SSCs were frozen at -80°C or -196°C. The results indicated that the optimal recovery rates of murine SSCs in the cryoprotective medium supplemented with LDL, trehalose and soybean lecithin were 92.53, 76.35 and 75.48% at -80°C, respectively. Compared with freezing at -196°C, the optimum temperature for improvement of recovery rates of frozen murine SSCs, cryopreservation in three different cryoprotectants at -80°C, were 17.11, 6.68 and 10.44% respectively. The recovery rates of murine SSCs in the cryoprotective medium supplemented with 0.2 M LDL were significantly higher than that of other cryoprotectants (P < 0.05). Moreover, the recovery rates were demonstrated to be greater at -80°C compared with at -196°C (P < 0.05). In conclusion, 0.2 M of LDL could significantly protect murine SSCs at -80°C. In the freezing-thawing process, LDL is responsible for the cryopreservation of murine SSCs because it can form a protective film at the surface of membranes. However, more research is needed to evaluate and understand the precise role of LDL during the freezing-thawing of SSCs.
精原干细胞（SSCs）具有自我更新能力，并为雄性生殖系的基因工程提供了一条途径。精原干细胞的冷冻保存对于治疗男性不育、精原细胞移植等具有潜在价值。为了研究不同冷冻保护剂对小鼠精原干细胞的冷冻保存效果，分别向冷冻保护培养基中添加0.2 M的低密度脂蛋白（LDL）、海藻糖和大豆卵磷脂，并将小鼠精原干细胞在-80°C或-196°C下冷冻。结果表明，在添加LDL、海藻糖和大豆卵磷脂的冷冻保护培养基中，小鼠精原干细胞在-80°C时的最佳回收率分别为92.53%、76.35%和75.48%。与在-196°C冷冻相比，在-80°C下用三种不同冷冻保护剂进行冷冻保存时，提高冷冻小鼠精原干细胞回收率的最佳温度分别为17.11%、6.68%和10.44%。添加0.2 M LDL的冷冻保护培养基中小鼠精原干细胞的回收率显著高于其他冷冻保护剂（P < 0.05）。此外，与在-196°C时相比，在-80°C时回收率更高（P < 0.05）。总之，0.2 M的LDL在-80°C时能显著保护小鼠精原干细胞。在冻融过程中，LDL负责小鼠精原干细胞的冷冻保存，因为它能在细胞膜表面形成保护膜。然而，需要更多的研究来评估和了解LDL在精原干细胞冻融过程中的精确作用。
