Pothana Lavanya, Devi Lalitha, Venna Naresh Kumar, Pentakota Niharika, Varma Vivek Phani, Jose Jedy, Goel Sandeep
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Animal House, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Cryobiology. 2016 Dec;73(3):356-366. doi: 10.1016/j.cryobiol.2016.09.169. Epub 2016 Sep 28.
Cryopreservation of immature testis is a feasible approach for germplasm preservation of male animals. Combinations of dimethyl sulfoxide (DMSO) and foetal bovine serum (FBS) are used for testis cryopreservation. However, an alternative to FBS is needed, because FBS is expensive. Buffalo ocular fluid (BuOF), a slaughter house by-product, could be an economical option. The objective of the present study was to assess whether BuOF can replace FBS for cryopreservation of immature mouse (Mus musculus), rat (Rattus norvegicus), and buffalo (Bubalus bubalis) testes. Results showed that rodent and buffalo testes frozen in DMSO (10% for rodents and 20% for buffalo) with 20% FBS or BuOF had similar numbers of viable and DNA-damaged cells (P > 0.05). The expression of cell proliferation- (PCNA) and apoptosis-specific proteins (Annexin V and BAX/BCL2 ratio) were also comparable in mouse and buffalo testes frozen in DMSO with FBS or BuOF (P > 0.05). Interestingly, rat testis frozen in DMSO with BuOF had lower expression of Annexin V protein than testis frozen in DMSO with FBS (P < 0.05). The percentage of meiotic germ cells (pachytene-stage spermatocytes) in xenografts from testis frozen either in DMSO with BuOF or FBS did not significantly differ in rats or buffalo (P > 0.05). These findings provide evidence that BuOF has potential to replace FBS for cryopreservation of immature rodent and buffalo testis. Further investigation is needed to explore whether BuOF can replace FBS for testis cryopreservation of other species.
未成熟睾丸的冷冻保存是雄性动物种质保存的一种可行方法。二甲基亚砜（DMSO）和胎牛血清（FBS）的组合用于睾丸冷冻保存。然而，由于FBS价格昂贵，需要一种替代物。水牛眼液（BuOF）是一种屠宰场副产品，可能是一种经济的选择。本研究的目的是评估BuOF是否可以替代FBS用于未成熟小鼠（小家鼠）、大鼠（褐家鼠）和水牛（水牛属）睾丸的冷冻保存。结果表明，用20% FBS或BuOF在DMSO中冷冻的啮齿动物和水牛睾丸，其活细胞和DNA受损细胞数量相似（P>0.05）。在DMSO中用FBS或BuOF冷冻的小鼠和水牛睾丸中，细胞增殖特异性蛋白（PCNA）和凋亡特异性蛋白（膜联蛋白V和BAX/BCL2比值）的表达也相当（P>0.05）。有趣的是，在DMSO中用BuOF冷冻的大鼠睾丸中，膜联蛋白V蛋白的表达低于在DMSO中用FBS冷冻的睾丸（P<0.05）。在DMSO中用BuOF或FBS冷冻的睾丸异种移植中，大鼠或水牛减数分裂生殖细胞（粗线期精母细胞）的百分比没有显著差异（P>0.05）。这些发现提供了证据，表明BuOF有潜力替代FBS用于未成熟啮齿动物和水牛睾丸的冷冻保存。需要进一步研究以探索BuOF是否可以替代FBS用于其他物种的睾丸冷冻保存。
