Proc. 9th CU. Vet. Sci. Ann. Con., 2010
115
The Effects of Culture Media Types and Embryo Density
on Development of Cat Embryos
1Faculty
T. Sananmuang*, T. Tharasanit, M. Techakumphu
of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand
*Corresponding author: [email protected]
Keywords: cat, culture media, developmental competence, embryo density
Introduction
A number of studies have demonstrated the
effects of culture media types (2, 3) and embryo
density (embryo numbers: media volume ratio)
(1) on embryo development. Knowledge on type
of culture medium and embryo density during
culture is very restricted in feline species. This
study aimed to determine the effects of culture
media types and embryo density on
developmental competence of cat embryos
cultured in vitro.
Materials and Methods
Immature cat cumulus oocyte complexes
(COCs) were matured and fertilized in vitro with
frozen-thawed semen. Approximately 8-10
embryos were randomly cultured in 20, 50 and
100 µl droplets (densitites: 1: 2, 1: 5 and 1: 10,
respectively) of three different culture media
(synthetic oviductal fluid: SOF, modified Earle’s
balanced salt solution: MK-1 and Tyrode’s
balanced salt solution: Tyrode). Cleavage,
morula and blastocyst rates were evaluated at
day 2, 5 and 7 of culture respectively. Embryonic
nuclei were assessed by staining the blastocysts
with DNA labeling (4', 6-Diamidino-2phenyindole: DAPI) and visualized with an
epifluorescent microscope (Olympus, Japan).
Results and Discussion
The results indicated that SOF (%
blastocyst: 61.0±7.7) and MK-1 (% blastocyst:
62.5±8.7) tended to support cat embryo
development better than Tyrode (% blastocyst:
33.3±9.8) (p<0.05) in 50 µl culture droplets.
Additionally, embryos cultured in 50 and 100 µl
droplets
tended
to
acquire
higher
developmental competence than those in 20 µl
culture droplets. Blastocyst rates of cat embryos
cultured in 50 and 100 µl droplets of SOF (%
blastocyst: 62.5±7.8 and 56.7±6.5) and MK-1 (%
blastocyst: 60.9±7.7 and 53.3±7.5) were similar,
and these two culture media supported
blastocyst formation better than Tyrode (%
blastocyst: 45.2±7.8 and 30.2±7.1, respectively).
Reducing
culture medium volume of SOF, MK-1and
Tyrode to 20 µl decreased blastocyst rates (%
blastocyst: 20.0±5.4, 39.6±6.8 and 13.6±5.2,
respectively). However, cell numbers of
blastocyst was not significantly affected by
different embryo culture media and volume.
Varied types of culture media and the
embryo density could affect the feline embryo
development differently. SOF and MK-1
supported embryo development better than
Tyrode. Embryos cultured in both 1:5 and 1:10
densities could develop better than those
cultured in 1: 20.
Acknowledgements
This work was supported by RGJ-PhD grant
and CHE-TRF Senior Research Fund (RTA
5080010).
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