Molecular Human Reproduction, Vol 4, 51-59, Copyright © 1998 by Oxford University Press
RT Pfaff, J Liu, D Gao, AT Peter, TK Li and JK Critser
Although embryo cryopreservation is routine for many mammalian species, it
is important to know how the fundamental cryobiology of these cells changes
with development. Progressive cleavage divisions result in a reduction in
the blastomere surface area available for water and cryoprotectant mass
transport. Therefore, the membrane permeability of murine oocytes, zygotes,
2-cell, 4-cell, and 8-cell embryos to water (Lp), and dimethylsulphoxide
(PDMSO), and the reflection coefficient, sigma (sigma) were determined.
Oocytes or zygotes were recovered, cumulus cells removed, then cultured
until use. Oocytes and embryos were immobilized and perfused with treatment
solutions at 24 degrees C. Osmotically induced cell volume changes over
time were videotaped followed by image analysis. The Lp values in the
presence of dimethylsulphoxide (DMSO) were 0.77, 0.81, 0.94, 0.86, and 1.10
microm/min/atm, and the PDMSO values were 1.85, 2.04, 2.41, 1.95, and
1.25x10(-3) cm/min for oocytes, zygotes, 2, 4, and 8-cell embryos
respectively. The Lp values in the presence of DMSO were significantly (P
< 0.05) higher than those in the absence of DMSO. Treating the whole
embryo as a single osmotic entity leads to significantly (P < 0.05)
elevated PDMSO estimates relative to those based upon measurements of
individual blastomeres. These data indicate that both Lp and PDMSO
estimates are lower when predicted on an individual blastomere basis. The
data also show that neither Lp nor PDMSO differ among oocytes, zygotes,
2-cell and 4-cell embryos. However, the significantly higher Lp and lower
PDMSO of the 8-cell stage support the hypothesis that fundamental
cryobiological differences may require developmental stage- specific embryo
cryopreservation protocols.
JOURNAL ARTICLE
Water and DMSO membrane permeability characteristics of in-vivo- and in- vitro-derived and cultured murine oocytes and embryos
Cryobiology Research Institute at Methodist Hospital of Indiana, Indianapolis 46202, USA.
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