In vitro Development of Preimplantation Caprine Embryo using Cryopreserved Black Bengal Buck Semen

Abstract

The aim of the present study was to produce goat embryo in different culture media through in vitro fertilization using cryopreserved black Bengal buck semen. Total 1265 fresh cumulus oocyte complexes (COCs) were collected by aspiration method with a 19 gauge hypodermic needle, washed 5-6 times and cultured in maturation media maintaining 5% CO2 level at 38.5ºC with maximum humidity in a incubator. After 27 h of incubation cumulus cells were stripped off from matured oocytes and transferred to acidified Tyrode’s medium for zona thinning and co-incubated with in vitro capacitated sperms for fertilization in Fert-BO media. In the experiment I, fresh buck semen and in experiment II, frozen buck semen was used for in vitro fertilization after in vitro processing. After 5 h of co-incubation, presumptive zygotes were washed and co-incubated with oviductal cells in three different culture media (RVCL, mSOF, KSOM) for further development. In fresh group cleavage (%) were 37.76 ± 2.98, 39.60 ± 1.75, 29.01 ± 1.74, and morula formation (%) were 7.72 ± 3.38, 6.03 ± 1.29, and 3.00 ± 3.00 in RVCL, mSOF and KSOM media respectively. However, in frozen group cleavage (%) were 29.17 ± 2.56, 27.70 ± 2.31, 24.17 ± 1.44 in RVCL, mSOF and KSOM media respectively and morula formation (%) was 2.93 ± 0.97 only in RVCL media. These results indicate that cryopreserved black Bengal buck semen have competence to produce embryos and could be used for embryo development in RVCL media through in vitro fertilization.