X-Message-Number: 14792
Date: Sun, 29 Oct 2000 11:22:05 -0800 (PST)
From: Doug Skrecky <>
Subject: vitrification solution VSv

Title
  Successful cryopreservation of mouse blastocysts using a new
  vitrification solution.
Source
  Journal of Reproduction & Fertility.  96(2):793-802, 1992 Nov.
Abstract
  Mouse blastocysts were exposed to solutions containing four concentrations
  (10, 20, 30 and 40% v/v) of six permeating cryoprotectants (glycerol,
  ethylene glycol, propylene glycol, dimethyl sulfoxide, 1,3-butanediol and
  2,3-butanediol) in phosphate-buffered saline (PBS) with calf serum (CS) at
  room temperature (20-22 degrees C). Blastocysts were exposed to these
  solutions for various periods, diluted into PBS plus CS with or without 1 mol
  trehalose l-1 solution and their subsequent survival in vitro was examined.
  Two-way anova showed a significant interaction (P < 0.01) between
  cryoprotectant type, concentration of cryoprotectant and method of dilution.
  However, no significant interaction was observed between cryoprotectant type
  and duration of exposure. Results suggest that cryoprotectant-induced injury
  to nonfrozen blastocysts is variable and depends on the cryoprotectant used.
  On the basis of toxicity assays, ethylene glycol was the
  least harmful and was combined with dimethyl sulfoxide and 1,3-butanediol to
  produce a new vitrification solution. Mouse blastocysts were
  successfully cryopreserved using a vitrification solution
  (designated as VSv) consisting of 20% ethylene glycol, 20% dimethyl sulfoxide
  and 10% 1,3-butanediol (v/v). Embryos were equilibrated in two steps, first
  in an equilibration solution (designated as ESv: 10% ethylene glycol, 10%
  dimethyl sulfoxide and 5% 1,3-butanediol; v/v) and then to VSv or one-step in
  VSv at different exposure times at room temperature, and then vitrified by
  direct plunging into liquid nitrogen. High developmental rates were obtained
  in vitro when the embryos were exposed to ESv and VSv for 3 and 0.5 min,
  respectively (96.2%) or exposed to VSv for 0.5 min (95.4%). Prolonged
  exposure time proved detrimental to subsequent embryo development in vitro.
  When vitrified warmed embryos were transferred immediately to pseudopregnant
  recipients, the rate of development to normal fetuses did not significantly
  differ from that of the nonvitrified control (two-step, 54.2 and one-step,
  45.0 versus 60.0%, P > 0.05). These results suggest that the simple
  vitrification solution described in this study is effective
  for the cryopreservation of mouse blastocysts.

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