牛精子染色体制备及影响因素探讨

Preparation of Bull Sperm Chromosome and Its Effect Factors

用去透明带金黄地鼠卵异种体外受精技术制备了牛精子单倍染色体。冻精复苏、洗涤、获能用 Ｈａｍ  Ｆ１０ 和 Ｒ Ｐ Ｍ Ｉ１６４０ 为主的混合培养基，辅以亚牛磺酸、肾上腺素，１０ μｍ ｏｌ／ Ｌ 钙离子载体（ Ｉ Ａ）处理牛精子 １２～１５ ｍ ｉｎ，诱导获能。异合卵用 Ｔ Ｃ１９９ 培养，以鬼臼毒素和长春花碱阻断原核融合和纺锤体形成，其浓度为常规浓度的 ２ 倍，在第 １ 次卵裂中期制备染色体。结果显示穿卵率达 ８３８％ ，可分析精子染色体核型成功率为 ４２４％ 。认为这是一种有效制备牛精子染色体的方法，并对其影响因素进行了探讨。

A method for preparation of bull sperm chromosome using interspecific in vitro fertilization with free zona golden hamster eggs was reported. The mixed medium of Ham F10 and RPMI 1640 supplemented with hypotaurine and epinephrine was used for handling bull sperm and hamster eggs. The thawed bull sperm was treated with Ca ionophore A23187 to induce capacitation. The fertilized eggs were cultured in medium TC199 and blocked to form karyogamy and spindle with high concentration of podophyllotoxin and vinblastine. Bull sperm chromosome was prepared in the first cleavage metaphase. The results showed that the rates of eggs penetration and analysable sperm karyotype were 83.8% and 42.4%, respectively. These results also demonstrated that this is an efficient method to prepare bull sperm chromosome. In addition, the effect factors of the experiment were discussed.