The present study investigated the

The present study investigated the effects of bovine granulosa cell monolayers (BGML) and canine granulosa cell monolayers
(CGML) on nuclear maturation of canine oocytes with and without cumulus cells. Cumulus-oocyte complexes (COCs) or
cumulus-free oocytes were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, control group), DMEM with BGML
(BGML group), or DMEM with CGML (CGML group) for 72 h at 38.5 °C in 5% CO2, 5% O2, and 90% N2. All media were
supplemented with 10% of FCS, 50 ng/mL of EGF, 2
g/mL of estradiol-17, 0.1 IU/mL of hCG, 0.1 IU/mL of FSH, 0.25 mM
of pyruvic acid, 100
M of -mercaptoethanol, 100 IU/mL of penicillin, and 100
g/mL of streptomycin. In cumulus-enclosed
oocytes retrieved from ovaries at estrus and/or diestrus, the highest percentage of M-II oocytes (P
0.05) was present in the
BGML group (27.0%) compared with the CGML group (7.9%) and the control group (3.5%). In cumulus-free oocytes collected
from ovaries at estrus and/or diestrus, the proportions of M-II oocytes co-cultured with the CGML were low (3.0%) and similar
(P  0.05) to proportions achieved with control (3.0%). However, the presence of BGML improved (P
0.05) the ability of
denuded oocytes to develop into M-II (10.2%). The BGML group had the highest overall meiotic resumption (P
0.05), and least
oocyte degeneration (P
0.05) among experimental groups. In conclusion, BGML had a positive impact on the in vitro
maturation system, as well as meiotic resumption of canine oocytes.
© 2012 Elsevier Inc. All rights reserved.

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