Human Reproduction, Vol 12, 2246-2250, Copyright © 1997 by Oxford University Press
H Van de Velde, ZP Nagy, H Joris, A De Vos and AC Van Steirteghem
The aim of this study was to compare concentrations of hyaluronidase and
mechanical methods used to denude human oocytes from surrounding cumulus
and corona cells prior to intracytoplasmic sperm injection (ICSI). Cumulus
and corona cells were removed in two pipetting steps: first in a medium
containing hyaluronidase, and then in a medium without enzyme. The first
step in the procedure was investigated. Different hyaluronidase
concentrations (78, 39 or 10 IU/ml) and pipettes of different size (inner
diameter 250 or > 1000 microm) were used to remove the cumulus cells.
The time required to denude the oocytes was recorded. Metaphase II oocytes
were injected, and the survival, fertilization, embryo development and
pregnancy rates were evaluated. We found that by using a pipette with an
inner diameter of at least 1000 microm we were able to decrease
significantly the time an oocyte is exposed to hyaluronidase, even if the
concentration of enzyme is very low (10 IU/ml). For the different
conditions there was no statistically significant effect on the outcome in
terms of survival, normal fertilization [two pronuclear (2PN)],
parthenogenetic activation (1PN), abnormal fertilization (3PN), embryo
development and pregnancy rates after ICSI. In conclusion, a concentration
as low as 10 IU/ml hyaluronidase in combination with a pipette of at least
1000 microm inner diameter can be used successfully to denude oocytes for
microinjection.
ARTICLES
Effects of different hyaluronidase concentrations and mechanical procedures for cumulus cell removal on the outcome of intracytoplasmic sperm injection
Centre for Reproductive Medicine, Dutch-speaking Brussels Free University (Vrije Universiteit Brussel), Belgium.
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