ISBN: 978-979-19096-1-7
Abstract
The oocyte-secreted polypeptide growth factors,
growth differentiation factor 9 (GDF-9) and bone
morphogenetic protein 15 (BMP-15, also known as
GDF-9B) have both been shown to be essential for
ovarian follicular growth and function. Bone
morphogenetic protein-15, an oocyte growth factor
belonging to the transforming growth factor-
superfamily, has recently been shown to be
necessary for normal female fertility in mammals.
In mice, the use of knock-out models has shown
that GDF-9 is essential for normal follicular
development with BMP-15 regulating the
fertilization potential of oocytes. The aim of this
research was to find out the influenced volume of
IVM oocyte media to BMP-15 and GDF-9 gene
expression. Oocytes were collected by puncturing
the surface of the mice ovaries with sterile needles
27 G . Furthermore oocytes were matured within
Hx medium during 22 hours at 38,5 C temperature
in the incubator with 5% CO2 to achive metaphase
II (M II) oocytes. Oocytes were cultured in 3
variations media droplet volume of 50 ul / droplet,
100 ul / droplet and 200 ul / droplet. After RT-PCR
and agarose gel electrophoresis, relative mRNA
abundance of BMP-15 and GDF-9 were analyze in
each group of oocytes. The BMP-15 gene
expression of oocytes were obtained when oocytes
were cultured in 50, 100 and 200 ul / droplet media.
The Highest expression of BMP-15 gene obtained
when oocytes were culture in 200 ul / droplet
media. The GDF-9 gene expression of oocytes were
obtained when oocytes were cultured in 200 ul /
droplet media.
Introduction
A. Rahayu, Growth Factors (BMP-15 AND GDF-9) Gene Expression Of Mice Oocytes In Vitro
Result
2 Methodology
Animals
Mice used in this study were maintained at the
Tohoku University animal house. Twenty-one-dold mice were injected with 5 IU PMSG, and
ovaries were collected 46 h later. All animals were
housed under controlled humidity, with a 12-hr
light and a2-hr dark phase, temperature, and fed ad
libitum.
Oocytes collecting
Mice were killed by the cervical dislocation.
Ovaries were cleaned free of adherent adipose and
connective tissues and placed in HEPES-buffered
tissue cultured medium-199 (H-TCM-199; ICN
Biomedicals Inc., Costa Mesa, CA) supplemented
with 0.1% (wt/vol) BSA (H-TCM-199/BSA).
COCs were isolated by puncturing antral follicles
with 27 G needles and collected in L-15Meiumd/BSA.
Only COCs with a uniform
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3rd International Conferences and Workshops on Basic and Applied Sciences 2010
ISBN: 978-979-19096-1-7
(a)
9 10 11 12
(b)
(c)
Figure 1: Follicle culture in vitro before (a) and
after (b) culturing oocytes. (c) MII Oocytes
obtained from in vitro culture
1 2
Conclusions
References
[1] Braw-Tel R & Yossefi S. 1997. Studies in
vivo and in vitro on the initiation of follicle
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A. Rahayu, Growth Factors (BMP-15 AND GDF-9) Gene Expression Of Mice Oocytes In Vitro
of
cultured
early
ovarian
Endocrinology 140 12361244.
follicles.
3rd International Conferences and Workshops on Basic and Applied Sciences 2010
B030
ISBN: 978-979-19096-1-7