Buarpung Sirirak, Tharasanit Theerawat, Thongkittidilok Chommanart, Comizzoli Pierre, Techakumphu Mongkol
Department of Obstetrics,Gynaecology and Reproduction,Faculty of Veterinary Science,Chulalongkorn University,Bangkok,Thailand.
Department of Reproductive Sciences,Center for Species Survival,Smithsonian Conservation Biology Institute,Washington,District of Columbia,USA.
Zygote. 2015 Oct;23(5):644-51. doi: 10.1017/S0967199414000276. Epub 2014 Jul 3.
The objective of this study was to compare the efficiency of preservation media for isolated feline testicular spermatozoa as well as the concentrations of bovine serum albumin (BSA) on: (1) the membrane (sperm membrane integrity (SMI)) and DNA integrity of spermatozoa; and (2) the developmental potential of spermatozoa after intracytoplasmic sperm injection (ICSI). Isolated cat spermatozoa were stored in HEPES-M199 medium (HM) or Dulbecco's phosphate-buffered saline (DPBS) at 4°C for up to 7 days. Results indicated that HM maintained a better SMI than DPBS throughout the storage periods (P > 0.05). When spermatozoa were stored in HM supplemented with BSA at different concentrations (4, 8 or 16 mg/ml), SMI obtained from HM containing 8 and 16 mg/ml BSA was higher than with 4 mg/ml BSA (P 0.05). In summary, cat spermatozoa immediately isolated from testicular tissue can be stored as a suspension in basic buffered medium at 4°C for up to 7 days. BSA supplementation into the medium improves membrane integrity of the spermatozoa during cold storage. Testicular spermatozoa stored in HM containing 16 mg/ml BSA retained full in vitro developmental potential after ICSI, similar to that of fresh controls even though DNA integrity had slightly declined.
本研究的目的是比较用于分离猫科动物睾丸精子的保存培养基的效率以及牛血清白蛋白（BSA）浓度对以下方面的影响：（1）精子的膜（精子膜完整性（SMI））和DNA完整性；（2）胞浆内精子注射（ICSI）后精子的发育潜能。将分离出的猫精子在4°C下于HEPES-M199培养基（HM）或杜氏磷酸盐缓冲盐水（DPBS）中保存长达7天。结果表明，在整个保存期内，HM维持的SMI比DPBS更好（P>0.05）。当精子在补充有不同浓度（4、8或16mg/ml）BSA的HM中保存时，从含有8mg/ml和16mg/ml BSA的HM中获得的SMI高于含有4mg/ml BSA的HM（P 0.05）。总之，从睾丸组织中立即分离出的猫精子可以作为悬浮液在4°C的基本缓冲培养基中保存长达7天。向培养基中添加BSA可改善精子在冷藏期间的膜完整性。即使DNA完整性略有下降，保存在含有16mg/ml BSA的HM中的睾丸精子在ICSI后仍保留了与新鲜对照相似的完全体外发育潜能。
