Effect of Somatic Cell Types and Culture Medium on in vitro
Maturation, Fertilization and Early Development Capability of
Buffalo
Oocytes
H. Jamil*, H. A. Samad, N. Rehman, Z. I. Qureshi and L. A. Lodhi
Department of Theriogenology,
University
of
Agriculture,
Faisalabad,
Pakistan
*Corresponding author:
drhjamil@hotmail.com
Abstract
This study was designed to evaluate the efficacy of different somatic cell types
and media in supporting in vitro
maturation (IVM), in vitro
fertilization (IVF) and early embryonic development competence of buffalo
follicular oocytes. Cumulus oocyte complexes were collected for maturation from
follicles (>6mm)
of buffalo ovaries collected at the local abattoir. Oocytes were co-cultured in
tissue culture medium (TCM-199) with either granulosa cells, cumulus cells, or
buffalo oviductal epithelial cells (BOEC) @ 3x106 cells/ml or in
TCM-199 without helper cells (control) at
39°C
and 5%CO2 in humidified air. Fresh semen was prepared in modified Ca++
free Tyrode medium. Fertilization was carried out in four types of media:
i) Tyrode lactate albumin pyruvate (TALP), ii) TALP+BOEC, iii) modified Ca++
free Tyrode and iv) modified Ca++ free Tyrode+BOEC. Fertilized
oocytes were cultured for early embryonic development in
TCM-199
with and without BOEC. Higher maturation rates were observed in the granulosa
(84.24%) and cumulus cells (83.44%) than BOEC co culture system (73.37%).
Highest fertilization rate was obtained in modified Ca++ free Tyrode
with BOEC co culture (70.42%), followed by modified Ca++ free Tyrode
alone (63.77%), TALP with BOEC (36.92%) and TALPalone (10.94%). Development of early embryos (8-cell stage) improved in
TCM-199
with BOEC co culture than
TCM-199
alone. From the results of this study, it can be concluded that addition of
somatic cells (granulosa cells, cumulus cells) results in higher maturation
rates of buffalo follicular oocytes than BOEC co culture system, while
fertilization rate improved in modified Ca++ free Tyrodewithand without BOEC. Addition
of BOEC to
TCM-199
improved the developmental capacity of early embryo.
Key words:
Buffalo
oocytes; In vitro maturation;
In vitro fertilization; Somatic cells
co culture