Wintenberger-Torres S, Popescu P C
Station centrale de Physiologie animale, I.N.R.A. 78350 Jouy-en-Josas, France.
Theriogenology. 1980 Nov;14(5):309-18. doi: 10.1016/0093-691x(80)90110-7.
We obtained 174 Charolais cow blastocysts at day 13 (D(13)) after superovulation and slaughter of 24 donor heifers. Using de Wecker's scissors, we sampled 1 10 to 1 3 of the total blastocyst volume. After colcemid treatment, hypotonic shock, fixation and Giemsa staining, the sex of 149 blastocysts (60 %) was determined. The main difficulty in sexing was the absence of metaphase or the presence of incomplete metaphases on the preparations. During manipulation, the portion of the blastocyst to be transferred was stored at 37 degrees C in a medium of 20 % fetal calf serum containing 80 % B(2) under an atmosphere of 90 % nitrogen, 5 % O(2) and 5 % CO(2) for 6 to 7 hours. After sex determination, the blastocysts were put into an artificial insemination straw and placed with a catheter at the base of the uterine horns. Twenty-eight blastocysts were transferred into both horns of 14 recipient Charolais heifers; 4 heifers (28 %) had an extended cycle and 3 (21 %) were pregnant. When 2, 3 or 4 blastocysts were transferred into the ipsilateral horn with the corpus luteum, 20 to 25 % of the recipients had an extended cycle and 37, 40 and 75 %, respectively, were pregnant. The number of surviving embryos per recipient never exceeded 2.
我们在对24头供体小母牛进行超排并于第13天(D(13))屠宰后,获得了174枚夏洛来牛囊胚。使用德韦克剪刀,我们采集了囊胚总体积的1/10至1/3。经秋水仙酰胺处理、低渗休克、固定及吉姆萨染色后,确定了149枚囊胚(60%)的性别。性别鉴定的主要困难在于制片上没有中期或存在不完全中期。操作过程中,待移植的囊胚部分在含有80%B(2)的20%胎牛血清培养基中,于90%氮气、5%氧气和5%二氧化碳的气氛下,在37℃保存6至7小时。性别鉴定后,将囊胚放入人工授精细管中,并用导管置于子宫角基部。将28枚囊胚移植到14头受体夏洛来小母牛的双侧子宫角;4头小母牛(28%)周期延长,3头(21%)怀孕。当将2、3或4枚囊胚移植到有黄体的同侧子宫角时,20%至25%的受体周期延长,分别有37%、40%和75%怀孕。每个受体存活胚胎的数量从未超过2个。