The effect of prolactin and bovine follicular fluid on the in vitro maturation and subsequent development of immature buffalo (Bubalus bubalis) oocytes

Sesay, A.R.; Shi DeShun

Journal of South China Agricultural University 25(2): 93-97

2004


ISSN/ISBN: 1001-411X
Accession: 004350559

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Abstract
Buffalo oocytes with compact cumulus cells were collected from slaughterhouse ovaries and cultured in TCM 199 supplemented with 5% oestrus cow serum (ECS) + 0.1 micro g/ml FSH and prolactin (PRL) 0, 0.1, 1.0, 10 micro g/ml in Experiment 1, and with 0% bovine follicular fluid (BFF) + 1.0 micro g ml PRL, 5% BFF, 5% BFF + 1.0 micro g/ml PRL in Experiment 2 and incubated at 38.5 degrees C and in phi =5% CO2 humidified air. After 24 to 26 h of maturation, IVF was done with swim-up separated frozen-thawed buffalo spermatozoa at 1x106/ml in modified Tyrodes medium (TALP). At 24 to 26 h post-insemination, the oocytes were co-cultured with granulosa cell monolayer in droplets containing the culture medium. The proportion of cleaved oocytes that became blastocysts within 9 days after co-culture with granulosa cell monolayer was then evaluated. In experiment 1, the proportion of cleaved oocytes that became blastocysts was higher (12.8%) at 1.0 micro g/ml PRL though not significantly different from the control (9.1%). In experiment 2, the addition of 5% BFF to the maturation medium significantly increased the cleavage rate of the oocytes when compared with the control (30.7 vs. 21.7%, P<0.05), but did not influence the proportion of cleaved oocytes that became blastocysts. The addition of 5% BFF + 1.0 micro g/ml PRL to the maturation medium had a cleavage rate of 38.1%, with 14.0% of the cleaved oocytes becoming blastocysts (P<0.05). It is concluded that the addition of appropriate amounts of prolactin and bovine follicular fluid to the maturation medium enhance the maturation of immature buffalo oocytes to become blastocysts.