X-Message-Number: 9449
Date: Sat, 11 Apr 1998 04:09:39 -0700 (PDT)
From: Doug Skrecky <>
Subject: ethylene glycol and glycerol

 Journal of Reproduction and Fertility 99: 471-477 1993
 J. Ali and J.N. Shelton
 "Design of Vitrification Solutions for the Cryopreservation of Embryos"

    A series of experiments was performed to determine the concentrations
 at which ten cryoprotectants singly and in pairs would vitrify on
 plunging into liquid nitrogen and remain vitreous when warmed by plunging
 into a water bath at 25C. From these tests eight solutions (VS) were
 selected for testing of toxicity to mouse morulae in vitro. One of these
 (VS1) was modified as a further five VS of which one (VS11) was tested
 for toxicity to all stages of mouse embroys and to sheep cpmpacted
 morulae. The concentrations at which the cryoprotectants vitrified on
 cooling were: butylene glycol, 3.0 mol/l; propylene glycol, 4.0 mol/l;
 dimethyl sulfoxide (DMSO) and glycerol 5.0 mol/l; ethylene glycol, 6.5
 mol/l. None of these, at the highest concentration tested, remained
 vitreous during warming. Methanol and the high molecular weight polymers,
 dextran, Ficoll, polyethylene glycol and polyvinylpyrrolidone, did not
 vitrify at the concentrations tested. Toxicity studies showed the order
 of increasing toxicity to be ethylene glycol, methanol, DMSO, glycerol,
 propylene glycol and butylene glycol. Of the mixtures composed of two
 cryoprotectants, those containing ethylene glycol and glycerol were the
 least toxic at vitrifying concentrations. VS11 (6.0 mol ethylene glycol/l
 and 1.8 mol glycerol/l) was well tolerated by mouse morulae, less well by
 eight- and one-cell embryos and poorly by two-cell embryos. Dilution of
 the VS11 from mouse embroys by exposure to 1.0 mol sucrose/l for 10 min
 did not enhance their survival. VS14 (5.5 ethylene glycol/l and 1.0 mol
 sucrose/l) was a good vitrifying mixture and was non-toxic to mouse
 embryos when they were exposed for up to 30 min. The survival of sheep
 compacted morulae in vitro was not affected by exposure to VS11 for up to
 20 min when dilution of the VS11 was done by exposure to 1.0 mol
 sucrose/l for 10 min. Without sucrose dilution, exposure to VS11 for 10
 min was detrimental to embryo survival.

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