Human Reproduction, Vol 13, 1595-1599, Copyright © 1998 by Oxford University Press
J Van der Elst, Y Amerijckx and A Van Steirteghem
We demonstrated previously that ultra-rapid freezing of mouse oocytes with
3.5 M dimethylsulphoxide (DMSO) decreased cell numbers in day 5 in- vitro
cultured blastocysts. In the present study we counted cell numbers of
trophectoderm (TE) and inner cell mass (ICM) separately following
differential labelling of TE with propidium iodide (red) and ICM with
bisbenzimide (blue). Blastocysts were from four groups of oocytes: (i)
cumulus-enclosed; (ii) hyaluronidase-treated cumulus-free; (iii)
cumulus-free and exposed to 3.5 M DMSO; and (iv) cumulus-free and
ultrarapidly frozen with 3.5 M DMSO. Mean (+/-SD) blastocyst cell numbers
were 54.7 +/- 22.0, 51.1 +/- 17.3, 52.3 +/- 13.1 and 40.4 +/- 18.4,
respectively. Mean TE cell numbers were 31.7 +/- 18.2, 28.9 +/- 13.3, 31.2
+/- 13.3 and 26.2 +/- 16.5 while mean ICM cell numbers were 23.0 +/- 9.4,
22.2 +/- 9.4, 21.1 +/- 7.3 and 14.2 +/- 7.3, respectively. Blastocyst and
ICM cell numbers were significantly lower in the group derived from
ultra-rapidly frozen oocytes compared with all other groups. Significantly
more blastocysts had < or = 32 cells and in blastocysts with > 64
cells a lower mean percentage of ICM was found. Ultra-rapid freezing of
mouse oocytes with 3.5 M DMSO can thus lead to day 5 in-vitro cultured
blastocysts with significantly decreased ICM cell numbers. The residual ICM
cell number in affected blastocysts may not reach a critical mass
sufficient for successful postimplantation development.
ARTICLES
Ultra-rapid freezing of mouse oocytes lowers the cell number in the inner cell mass of 5 day old in-vitro cultured blastocysts
Centre for Reproductive Medicine, Brussels Free University Hospital, Belgium.
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