Volume 7, Issue 2 (March 2005) 7, 139–144; 10.1111/j.1745-7262.2005.00039.x
Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender
Ya-Hui Li, Ke-Jun Cai, Lei Su, Mo Guan, Xie-Chao He, Hong Wang, Andras Kovacs and Wei-Zhi Ji
1.Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China
2.Graduate School of the Chinese Academy of Sciences, Beijing 100039, China
3.College of Food Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, China
4.Research Institute for Animal Breeding and Nutrition, 2053 Herceghalom, Hungary
Received: 2004-07-22 Accepted: 2005-02-04
Aim: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender.
Methods: Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris (mTTE) synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1%, 3%, 5%, 10% and 15% [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity.
Results: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P<0.05) for 5 % glycerol (42.95±2.55 and 50.39±2.42, respectively) than those of the other groups (1 %: 19.19±3.22 and 24.84±3.64; 3 %: 34.23±3.43 and 41.37±3.42; 10 %: 15.68±2.36 and 21.39±3.14; 15 %: 7.47±1.44 and 12.90±2.18). The parameters for 30 min equilibration (42.95±2.55 and 50.39±2.42) were better (P<0.05) than those of the other groups (10 min: 31.33 3.06 and 38.98± 3.31; 60 min: 32.49±3.86 and 40.01±4.18; 90 min: 31.16±3.66 and 38.30±3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity.
Conclusion: Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.
Keywords: cryopreservation, Macaca fascicularis, chemically defined extender, glycerol
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