Human Reproduction, Vol 13, 971-978, Copyright © 1998 by Oxford University Press
A Cooper, SJ Paynter, BJ Fuller and RW Shaw
The aim of this study was to develop a maturation protocol for immature
oocytes and assess the protocol with cryopreserved oocytes. Nuclear
maturation (mature spindle and aligned chromosomes) occurred irrespective
of the treatment regime: 71-89% of oocytes matured in vitro had a normal
spindle and chromosomes compared with 87% matured in vivo. Fertilization
rates were not significantly different from those of in-vivo matured
oocytes. Of the maturation treatment regimes investigated, the initial
treatment producing best development to blastocyst (cytoplasmic maturation)
involved a 2 h incubation in standard maturation medium (SMM) containing
7.5 IU follicle stimulating hormone (FSH) followed by 14 h in SMM plus 7.5
IU FSH:luteinizing hormone with follicular cells [62% (range 49-69)]. The
addition of 1 ng/ml epidermal growth factor (EGF) in this protocol resulted
in development [75% (range 71-81)] that was not significantly different
from in-vivo matured oocytes [82% (range 73-90)]. Exposure of the oocytes
to 1.5 M dimethylsulphoxide (DMSO) did not affect fertilization or
development rates. Following a slow-cool/thaw freezing regime, 81% (range
74-89) of the oocytes were morphologically normal, i.e. had a spherical
shape with an intact zona and oolemma; they had, however, lost their
previously attached cumulus and corona cells. Maturation of frozen-thawed
oocytes in the presence of EGF gave good fertilization rates but poor
development rates [80% (range 77-86) and 37% (range 33- 40) respectively].
In conclusion, the best maturation, both nuclear and cytoplasmic, occurred
in the presence of a combination of gonadotrophins, EGF and follicular
cells. Oocytes cryopreserved using a slow-cool/thaw regime can be matured
to produce blastocysts after in- vitro fertilization.
ARTICLES
Differential effects of cryopreservation on nuclear or cytoplasmic maturation in vitro in immature mouse oocytes from stimulated ovaries
Department of Obstetrics and Gynaecology, University of Wales College of Medicine, Heath Park, Cardiff, UK.
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