Enhancement of Developmental Competence of Immature Cattle Oocytes with Leukemia inhibitory Factor as a Culture Media Supplement

Abstract

Background: Leukemia Inhibitory Factor (LIF) plays an essential role in oocyte maturation and during early embryonic development. The present study aims to improve the in vitro cattle embryo production by supplementing culture media with LIF. Methods: Fresh ovaries and oviducts were collected from an abattoir in 0.9% saline (30-35°C) supplemented with antibiotics. Total 542 cumulus-oocyte complexes were aspirated from ovaries and cultured in maturation media in 5% CO2 incubator at 38.5°C with maximum humidity after 5-6 times washing. After 24 h matured oocytes were co-incubated with in vitro capacitated sperms in FBO medium. After 15-18 h cumulus cells were stripped off and presumptive zygotes were cultured in mCR2aa medium. After 40 to 42 h, cleavage was observed and embryos were cultured for 7-9 days. Culture media used to replace with fresh media after every 24 h. LIF was supplemented with 15, 30, 45 ng/ml. Result: Supplementation of LIF in culture media increased maturation rate, cleavage rate significantly (P less than 0.05). LIF Supplementation @ 30 ng/ml during culture increased blastocyst development (in control group 7.0±1.3 and 7.1±1.2, 12.9±0.4, 10.2±1.5 in 15 ng/ml, 30 ng/ml and 45 ng/ml respectively) significantly.