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当前位置: 首页 > 文章 > 绵羊冷冻胚胎移植成功 畜牧兽医学报 1982,13 (1) 17-23
Position: Home > Articles > SUCCESSFUL TRANSFER OF FROZEN EMBRYOS IN SHEEP Chinese Journal of Animal and Veterinary Sciences 1982,13 (1) 17-23

绵羊冷冻胚胎移植成功

作  者：
朱裕鼎;朱成宽;郭年藩;蒋世娥;罗应荣;冯书堂
摘  要：
以成年寒羊母羊做供、受体。使用 FSH(300单位)和 LH(150单位),进行供体超数排卵,并在发情周期的第6～8天(发情开始当天=0天)用杜氏磷酸盐缓冲液(PBS)+5%失活犊牛血清,冲洗子宫角手术采卵。只有形态正常的胚胎用于冷冻。胚胎冻前分6步(0.25M、0.5M、0.75M、1.0M、1.25M 和1.5M,每种浓度停留7～10分钟)添加抗冻剂(DMSO)。解冻后也分6步(1.5M、1.25M、1.0M、0.75M 和0.25M,每种浓度停留10分钟)除去 PBS 液中的 DMSO。胚胎在盛有0.25～0.3毫升 PBS 液(Whitting-ham,1971)的安瓿(容量1.0毫升)中冷冻,PBS 液含1.5M 的 DMSO。自1979年8月24日至12月27日,胚胎的冷冻和移植试验分三批进行。第一批胚胎是按 Willadsen(1976)冷冻羊胚胎的方法冷冻的,并降温至-120℃时将安瓿移入液氮。安瓶从-80℃--5℃,以12℃/分钟的速度解冻,从-5℃放入手中快速升温解冻。第二和第三批,胚胎是按 Willadsen(1978)所述及的牛胚胎冷冻和解冻方法进行的,并降温至-60℃将安瓿直接投入液氮。将安瓶放入-50℃的酒精浴中,以4℃/分钟的速度解冻至-10℃。从-10℃将安瓿放入手中快速解冻。第三批试验的受体根据母羊的健康和繁殖情况进行了挑选。共49个胚胎(47个形态正常的和2个形态异常的)移入23只受体,4只母羊妊娠。其中一只受体早产,羔羊死亡;一只受体母羊产了3只羊羔,共产了五只羊羔。三批试验的受体妊娠率和胚胎成活率分别如下:0%(0/8)、0%(0/18)、11.1%(1/9)、6.2%(1/16)和50%(3/6)、33.3%(5/15)。
译  名：
SUCCESSFUL TRANSFER OF FROZEN EMBRYOS IN SHEEP
作  者：
Zhu Yü-ding Zhu Cheng-kuan Guo Nian-fan Jiang Shi-e Luo Ying-rong Feng Shu-tang (Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing)
摘  要：
Mature Hu-Young ewes were used as donors and recipients.Supe- rovulation was induced with FSH(300u.)and LH(150u.),and the em- bryos were collected surgically from the uterine horns on Day 6-8(on- set of oestrus=Day o)by flushing with Dulbecco's phosphate-buffered saline(PBS)+5% heated calf serum. Only morphologically normal embryos were used for freezing.Befo- re freezing additiou of cryoprotectant(DMSO)was carried out in six steps(0.25M,0.5M,0.75M,1.0M,1.25M and 1.5M for 7-10 min each). After thawing the DMSO in PBS was removed also in six steps(1.sM, 1.25M,1.0M,0.75M,0.5M and 0.25M for 10 min in each concentration of PBS),Embryos were deep-frozen in ampoules(volume 1.0ml)contai- ning 0.25—0.3ml of 1.5 M—DMSO in PBS(Whittingham,1971). The experiments on the freezing of embryos,from 24 August to 27 December 1979,were divided into three trials.In trial Ⅰ the embryos were frozen according to the method of S.M.Willadsen(1976)in sheep, and from-120℃ the ampoules were thawed from -80℃ to -5℃ at 12℃/min,and from -5℃ the ampoules were thawed rapidly in hand.In trials Ⅱ and Ⅲ the embryos were frozen and thawed as descri-bed by S.M.Willadsen (1978) in cow,and from -60℃ the ampoules were plunged directly into liquid nitrogen.The ampoules were transfer-red to an ethanol bath at -50℃ and thawed at 4℃/min t0-10℃.F-rin -10℃ the ampoules were reroarmed rapidly in hand.A total of 49 embryos (47 morphologically normal and 2 abnormal) were transferred to 23 recipients.Four ewes were pregnant,One of them casted a dead lamb.Five lambs were born,and three of them were from one recipient ewe.Pregnant rates of recipients and survival rates of embryos in the 3 trials were as follows respeetively 0%(0/8),0%(0//8);11.1%(1/9),6.2(1/16) and 50%(3/6),33.3%(5/15).

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