ScienceAsia 15 (1989): 251-260 |doi: 10.2306/scienceasia1513-1874.1989.15.251

CO-CULTURE OF BOVINE EMBRYOS FROM OOCYTES MATURED AND FERTILIZED in vitro TO THE BLASTOCYST STAGE WITH OVIDUCTAL TISSUE*

YINDEE KITIYANANT ,^a CHAROENSRI THONABULSOMBAT ,^a CHAINARONG TOCHARUS ,^a BOONYAWAT SANITWONGSE^b AND KANOK PAVASUTHIPAISIT^a

ABSTRACT: The development of in vitro matured (IVM), in vitro fertilized (IVF) bovine embryos in co-culture with oviductal tissue suspension was investigated. Follicular oocytes (n =2929) collected from cattle at local slaughter houses were matured and fertilized in vitro. The follicular oocytes were graded according to their investments, Le. compact, expanded, partially denuded and completely denuded as grades 1,2,3, and 4, respectively. Of 1134 ova in grade 1, 966 (85%) were penetrated by spermatozoa, fertilized and cleaved to a 2-ceIl stage at 30-48 h post insemination. In oocyte grades 2,3 and 4, 72%, 64% and 63% were observed respectively to be 2-ceIl. Bovine oviductal tissue culture was prepared by scraping luminal tissue of intact oviduct with a glass slide and suspended in TALP+ 10% heat-treated fetal calf serum. Zygotes were placed in treatment culture 18-22 h after insemination. Cleavage of the embryos up to the 6- to 8- cell stage were possible in all groups of oocytes. However, the development to late morula (LM) or blastocyst (BL) was observed only in the oocytes of grades 1 and 2. These results demonstrate that immature oocytes with cumulus cells can be matured and fertilized in vitro, and oviductal tissue co-culture can support normal development of I VM-IVF bovine embryo to LM and BL without using any intermediate recipients.

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^a) Department of Anatomy, Faculty of Science, and Institute of Science and Technology for Research and Development, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand.
^b) Artificial Insemination Center, Livestock Division, Pathumthanee, Thailand.

Received 6 October 1989