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KMID : 0900919920160030239
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Korean journal of Animal Reproduction
1992 Volume.16 No. 3 p.239 ~ p.246
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In Vitro Fertilization of Bovine Oocytes Matured In Vitro by Microinjection of Spermatozoa
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Kim Seon-Ku
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Abstract
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Capacitated and acrosome~reacted spermatozoa were microinjected into the perivitelline space of bovine oocytes matured in vitro. Oocytes obtained from the ovaries of slaughtered heifers and cows were cultured in vitro in the TCM-199 supplemented with 20% FCS for 24 hr at 39^{circ}C under an atmosphere of 5% CO_2 8% O_2. Fresh or frozen spermatozoa were incubated for 2 hr at 39°C under an atmos-phere of 5% CO_2, 8% O_2 in Ham’s F-lO medium containing 0.75% BSA for capacitation, and kept for 30 min in culture medium containing 12 mM of dbcGMP and lOmM of immidazol for acrosome resction. One motile spermatozoon was injected into the perivitelline space of each oocyte. The 2nd polar body and the pronuclei were observed in 9.5% and 5.4% of oocytes, respectively. The rate of cleavage of oocyte over 2-cell stage was 4.1%(10 of 242), These results indicate that the microinjection may be a useful technique to study sperm-oocyte interaction.
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KEYWORD
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bovine oocytes, microinjection of spermatozoa, in vitro fertilization
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