REPRODUCTION - Facultad de Ciencias Veterinarias

Abstract
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REPRODUCTION
Oxidative stress is implicated in loss of embryos through apoptosis. The results of study demonstrates that proportion of
embryos exhibiting apoptotic morphology is more at advanced stages of development with relative abundance of Bax is
higher than Bcl XL and cysteamine supplementation did not have significant effect on the apoptosis and relative
abundance of Bax and Bcl XL.
INTRODUCTION
Apoptosis and effect of cysteamine
supplementation in IVM and IVC media
during in vitro development of buffalo
(Bubalus bubalis) embryos
The process of in vitro embryo production faced many changes since the birth of first in vitro produced (IVP) buffalo
calf; however the final yield of blastocysts is low. Embryos in in vitro development are vulnerable to oxidative stress
thereby the in vitro embryos suffer supra physiological oxygen tension and toxicity elicited by ROS. Supplementation of
thiols such as cysteamine and Beta mercaptoethanol in embryo culture medium protected embryos from oxidative stress
and improved the production efficiency 1,2,3 . The aim of the present study was to investigate the process of apoptosis in
buffalo in vitro produced embryos and the effect of cysteamine on the development of in vitro buffalo embryos with
respect to apoptosis.
MATERIALS AND METHODS
Anand V 1 ; Singh KP 1 ; Palta P 1 ; Manik RS 1 ; Singla S K 1 ; Chauhan MS 1,2
1 Animal Biotechnology Centre, National Dairy Research Institute,
karnal-132001, INDIA
E- mail: chauhan_abtc@gmail.com
Production of in vitro fertilized buffalo embryos
Keywords: apoptosis - cysteamine - In Vitro - buffalo
In vitro production of buffalo embryos was done as described in 1 .For experimental purpose in vitro maturation and in
vitro cultre media was supplemented with cysteamine at 50ìM and 100 ìM respectively.
Assessment of apoptosis by Acridine orange and Ethidium bromide (AO/EB staining)
Assessment of apoptosis in different stages of embryos wad done as described in 4 . Embryos treated with 100 ìM H2O2 was
used as control. Embryos are examined under fluorescence microscope (Excitation wavelength 450-490 nm; barrier filter
515nm) after washing them in culture medium.
Proceedings 9 th World Buffalo Congress