Vitrification properties of solutions of ethylene glycol in saline containing PVP, Ficoll, or dextran.
Vitrification solutions which are used for cells or embryos generally contain cryoprotectant, physiological saline, and one or more macromolecular solutes. The macromolecules modify the vitrification tendencies of these solutions, but there is little detailed information on the vitrification properties of ethylene glycol solutions containing the additives PVP, Ficoll, and dextran. This study therefore added ethylene glycol to 0.9% NaCl in water (saline) and used differential scanning calorimetry to determine the lowest concentration at which the solution would remain vitreous when a warming rate of 10 degrees C/min was used. In the absence of other additives 59 wt% ethylene glycol (EG) in saline formed a stable glass. When ethylene glycol was replaced by the polymers Ficoll and/or dextran on a weight for weight basis, the resulting solution vitrified less readily than an EG-saline solution even though the total solute concentration was kept constant. The total solute concentration required to form a stable vitreous solution increased as the Ficoll 70,000 and 400,000 MW or dextran 78,000 MW content increased (5, 10, and 20 wt%). Ficoll and dextran had little or no effect on the glass transition and melting points of the solutions. In the presence of PVP vitrification occurred at a total solute concentration of 59 wt% (PVP 360,000 MW) or 60 wt% (PVP 40,000 MW) for all three tested PVP concentrations (5, 10, and 20 wt%). Although this indicates that PVP and EG have comparable vitrification properties, the melting and the glass transition temperature of the solutions rose as the PVP content increased. When 1 m sucrose was added to saline and 0, 5, 10, or 20 wt% PVP 40,000 MW vitrification was achieved with 31, 26, 23, and 15% EG, respectively, indicating that the total solute concentration required for vitrification could be estimated with reasonable accuracy from the sum of the individual components. We conclude that the tested polymers differ in how they interact with ethylene glycol-based vitrification solutions.
Shaw
J M
JM
Institute of Reproduction and Development, Monash University, Clayton, Victoria 3168, Australia.
Kuleshova
L L
LL
MacFarlane
D R
DR
Trounson
A O
AO
D002152:Calorimetry, Differential Scanning; D003451:Cryoprotective Agents / Q000737:chemistry; D003911:Dextrans / Q000737:chemistry; D019855:Ethylene Glycol; D005069:Evaluation Studies as Topic; D005362:Ficoll / Q000737:chemistry; D008970:Molecular Weight; D011205:Povidone / Q000737:chemistry; D012965:Sodium Chloride; D012996:Solutions; D013816:Thermodynamics
D003451:Cryoprotective Agents; D003911:Dextrans; D012996:Solutions; D005362:Ficoll; D012965:Sodium Chloride; D019855:Ethylene Glycol; D011205:Povidone