To optimize In vitro maturation (IVM) of quality oocytes for embryo production through IVF and SCNT.Methods
Buffalo oocytes were in vitro matured in the presence of the pokeweed lectin (Phytolacca americana), a potent lymphocyte mitogen. Lectin was supplemented in TCM + 10% FBS at the doses of 0, 1, 5, 10, 15, 20 and 40 μg/ml and cumulus expansion and gene expression patterns were characterized.Results
The degree of cumulus expansion in different lectin treatment levels improved from 1.1 at 1 Ag/ml level to 3.60 at 10 μg/ml level and then decreased in higher concentration 20 μg/ml (1.66) and 40 μg/ml (0.64). IVF embryos showed highest cleavage rate (88.8%) in 10 μg/ml lectin treatment. Expression of all mRNA transcript studied (Cx43, GDF 9, FGF-4 and Fibronectin) was positively correlated with cumulus expansion and polar body extrusion.Conclusions
Mitogenic lectin supplemented maturation media improves oocyte quality for in vitro embryo production.