淘汰母梅花鹿再利用模式的初探:体内外胚胎的生产

Exploration of a reuse pattern of culled female Sika deer:In vivo and in vitro embryo production

试验旨在探索体内胚胎生产+活体采卵+离体取卵模式在淘汰母鹿再利用中的可行性。试验以4只淘汰母梅花鹿作为研究对象,先用羊用阴道内孕酮释放装置(CIDR)+促卵泡激素(FSH)+孕马血清促性腺激素(PMSG)方案进行超排,生产体内胚胎。冲胚结束时,对供体鹿两侧卵巢进行手术法吸卵,获取体内卵母细胞。最后随机选择2只母鹿屠宰获取体外卵母细胞,并利用所获卵母细胞研究DSOF液体系和SOF液体系在梅花鹿体外胚胎生产中的使用效果。结果显示,经超排处理后,所有供体母鹿均出现了发情表现,并有3只配种成功,平均冲胚2.7枚/只,回收率33.3%,平均获可移植胚0.3枚/只,可移植胚率12.5%。对供体鹿进行活体采卵共获卵母细胞6枚,回收率26.1%,仅1枚卵母细胞可用,可用率16.7%。对2只母鹿进行离体取卵共获可用卵丘-卵母细胞复合体(COCs)14枚,将该14枚COCs分别于DSOF液体系、SOFaa+E-DSOF和SOF液体系中受精与培养,卵裂率分别为22.2%(2/9)、50.0%(1/2)和66.7%(2/3)。仅SOF液体系中有一个受精卵发育到了8-cell阶段,其余所有卵裂胚均停滞在2-cell阶段。研究表明,体内胚胎生产+活体采卵+离体取卵模式可用于淘汰母鹿的再利用,但试验中使用技术的效率还有待进一步提高。SOF液体系可用于我国梅花鹿至少8-cell体外胚胎的生产。

The experiment aimed to explore the feasibility of the in vivo embryo production+ovum pick-up+in vitro oocyte retrieval model in the reuse of culled female deer.Four culled female Sika deer were used as research subjects.A controlled internal drug release(CIDR)combined with follicle-stimulating hormone(FSH)and pregnant mare serum gonadotropin(PMSG)protocol was first used for superovulation to produce in vivo embryos.At the end of the flushing process,the ovaries of the donor deer were surgically aspirated to obtain oocytes.Finally,two female deer were randomly selected for slaughter to retrieve in vitro oocytes,and the obtained oocytes were used to study the effectiveness of the DSOF liquid system and SOF liquid system in the ex vivo embryo production of Sika deer.The results showed that after superovulation treatment,all donor deer exhibited estrous behavior,and three of them were successfully mated,with an average of 2.7 flushed embryos per deer,a retrieval rate of 33.3%,an average of 0.3 transferable embryos per deer,and a transferable embryo rate of 12.5%.A total of six oocytes were obtained from the live oocyte recovery of the donor deer,with a retrieval rate of 26.1%,and only one oocyte was usable,with a usability rate of 16.7%.A total of 14 usable cumulus-oocyte complexes(COCs)were obtained from the in vitro oocyte retrieval of two female deer.These 14 COCs were fertilized and cultured in the DSOF liquid system,SOFaa+E-DSOF,and SOF liquid systems,with cleavage rates of 22.2%(2/9),50.0%(1/2),and 66.7%(2/3),respectively.Only one fertilized egg in the SOF liquid system developed to the 8-cell stage,while all other cleavage embryos were arrested at the 2-cell stage.The study indicates that the in vivo embryo production+ovum pick-up+in vitro oocyte retrieval model can be used for the reuse of culled female deer,but the efficiency of the techniques used in the experiment still needs to be further improved.The SOF liquid system can be used for the production of at least 8-cell in vitro embryos in Chinese sika deer.