Improvement of in vitro oocyte maturation with lectin supplementation and expression analysis of Cx43, GDF-9, FGF-4 and Fibronectin mRNA transcripts in Buffalo (Bubalus bubalis)

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Abstract

Purpose

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.

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