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源自体外成熟、受精并与牛输卵管上皮细胞共培养的卵母细胞的牛胚胎的发育与活力

Development and viability of bovine embryos derived from oocytes matured and fertilized in vitro and co-cultured with bovine oviducal epithelial cells.

作者信息

Xu K P, Yadav B R, Rorie R W, Plante L, Betteridge K J, King W A

机构信息

Department of Biomedical Sciences, University of Guelph, Ontario, Canada.

出版信息

J Reprod Fertil. 1992 Jan;94(1):33-43. doi: 10.1530/jrf.0.0940033.

Abstract

A co-culture system using a suspension of detached bovine oviducal epithelial cells (BOEC) has been developed as an effective culture method for supporting the development of bovine embryos derived from oocytes matured and fertilized in vitro. Four commercially available culture media (Waymouth's, Ham's F-10, TCM 199 and Ménézo's B2) supplemented with 10% oestrous cow serum, and a modified Tyrode's medium (TALP) supplemented with 0.6% bovine serum albumin were used. Ménézo's B2 resulted in the highest percentages of total uncleaved presumptive zygotes, and of the cleaved zygotes that reached at least the morula stage (31-46% and 66-74%, respectively). The embryos produced in vitro in B2 with BOEC resembled embryos produced in vivo with regard to numbers of cells (averaging 45.4 in morulae, 101.5 in blastocysts, 174.7 in hatching blastocysts and 195.9 in hatched blastocysts), rate of development (hatching on Day 8-9 of culture in vitro), rate of hatching (66% of cleaved zygotes) and pregnancy rates (63%) resulting from the transcervical transfer of selected embryos.

摘要

一种使用分离的牛输卵管上皮细胞(BOEC)悬液的共培养系统已被开发出来,作为一种有效的培养方法,用于支持源自体外成熟和受精卵母细胞的牛胚胎的发育。使用了四种市售培养基(韦茅斯培养基、哈姆氏F-10培养基、TCM 199培养基和梅内佐氏B2培养基),添加10%发情母牛血清,以及一种添加0.6%牛血清白蛋白的改良泰勒氏培养基(TALP)。梅内佐氏B2培养基导致未分裂的假定受精卵总数以及至少发育到桑椹胚阶段的分裂受精卵百分比最高(分别为31%-46%和66%-74%)。在B2培养基中与BOEC一起体外产生的胚胎在细胞数量(桑椹胚平均为45.4个,囊胚为101.5个,孵化囊胚为174.7个,孵化后囊胚为195.9个)、发育率(体外培养第8-9天孵化)、孵化率(分裂受精卵的66%)以及所选胚胎经宫颈移植后的妊娠率(63%)方面与体内产生的胚胎相似。

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