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In Vitro Maturation, Fertilization, Embryo Culture And Activation Treatments Of Ovine Oocytes In Different Culture Systems

KrishiKosh

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Title In Vitro Maturation, Fertilization, Embryo Culture And Activation Treatments Of Ovine Oocytes In Different Culture Systems
 
Creator Kundu, Akshaya Kumar
 
Contributor Thangavel, A
Jayaprasad, I. Alfred
Arunachalam, S
Palanisamy, A
 
Subject IVM
IVF
IVC
Ovine oocytes
Parthenogenic embryos
 
Description There is a rapid development in technologies to produce large number of
mammalian embryos, in vitro at low cost for research and commercial purposes,
still then in vitro developmental capacity of ovine oocytes is significantly lower
than that of in vivo system. Generation of a lot of data may be useful to optimize
the requirements for oocyte development, in vitro. This study was aimed to
improve ovine oocyte in vitro maturation, fertilization, embryos development and
activation of the ovine oocytes to produce parthenogenic embryos.
The cumulus oocyte complexes (COCs) were harvested by slicing method
from slaughterhouse-derived sheep ovaries. The COCs were matured in different
in vitro culture systems, viz., 1. maturation of oocytes in TCM-199, SOF and
Ham's F-10 media, 2. supplementation of IGF-I (100 ng/ml), EGF (10 ng/ml) and
IGF-I (100 ng/ml) + EGF (10 ng/ml), 3. supplementation of cysteamine @ 50,
100, 200 m and 4. supplementation of ß-mercaptoethanol (ß-ME) (12.5, 25, 50M) to the oocyte maturation medium. The cauda epididymal sperm from
slaughterhouse-derived testes were utilized for in vitro insemination of the
oocytes. The oocytes and sperms were co-incubated over the BRL cell and
OOEC monolayers in the fert-TALP medium with and without heparin. The
ovine embryos were cultured in different culture systems viz: 1. embryo culture
in TCM-199, SOF and Ham's F-10 media, 2. supplementation of IGF-I (100
ng/ml), EGF (10 ng/ml) and IGF-I (100 ng/ml) + EGF (10 ng/ml) to the IVC
medium, 3. embryo co-culture with BRL, Vero, BRL/Vero mixed cell and OOEC
monolayers and 4. supplementation of Hb (1 g/ml), L-NAME (1 M) and Hb
(1 g/ml) + L-NAME (1M) to the IVC medium. The matured ovine oocytes
were activated by ethanol, calcium ionophore and thimerosal to produce
parthenogenic embryos. The results were observed as follows.
TCM-199 and SOF media were found suitable for in vitro ovine oocyte
development. IGF-I, EGF and their combination accelerated the oocyte
maturation leading to increased fertilization rate and improved embryo
development. The presence of cysteamine and ß-ME during IVM were found
beneficial for ovine oocyte development. The presence of BRL cell and OOEC
even in the absence of heparin significantly improved the fertilization and embryo
development suggesting their capacitation and sperm viable ability. The SOF
medium showed better competence for ovine embryo culture over TCM-199 and
Ham's F-10 media. IGF-I, EGF and their combination in IVC medium enhanced
the embryo development rate and increased cell number in blastocysts. BRL,
Vero, BRL/Vero mixed cell and OOEC monolayers significantly improved the in
vitro embryo development. The activated parthenogenic embryos produced were
found to be quantitative but not qualitative compared to the IVF oocytes.
 
Date 2016-05-27T11:42:18Z
2016-05-27T11:42:18Z
2003
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/66365
 
Language en
 
Format application/pdf
 
Publisher Tamil Nadu Veterinary and Animal Sciences University