Cryopreservation of biological materials in a non-frozen or vitreous state
First Claim
1. An improved method for the cryopreservation of biological materials wherein the biological material is cooled to a vitreous state under pressure in the presence of an aqueous vitrification solution containing penetrating glass-forming solutes, the improvement comprising (a) replacing a sufficient amount of said penetrating solute in said solution with an equivalent amount of non-penetrating glass-forming solute to reduce damage to the biological material when exposed thereto;
- (b) treating the biological material with the solution obtained from step (a) at a rate and duration sufficient to prevent cells in said biological material from returning to isotonic volume prior to vitrification; and
(c) then subjecting the biological material to a pressure sufficient for vitrification upon cooling without substantial nucleation or ice crystal growth and without significant injury to the biological material.
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Abstract
A method is provided for the successful cryopreservation of biological materials including whole organs, organ sections, tissues and cells, in a non-frozen (vitreous) state, comprising cooling the biological material to be preserved under pressure in the presence of a non-toxic vitrifable protective solution to at least the glass transition temperature thereof to vitrify the solution without substantial nucleation or ice crystal growth and without significant injury to the biomaterial. The invention also provides non-toxic protective vitrification solutions useful in the cryopreservation of biomaterials.
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Citations
18 Claims
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1. An improved method for the cryopreservation of biological materials wherein the biological material is cooled to a vitreous state under pressure in the presence of an aqueous vitrification solution containing penetrating glass-forming solutes, the improvement comprising (a) replacing a sufficient amount of said penetrating solute in said solution with an equivalent amount of non-penetrating glass-forming solute to reduce damage to the biological material when exposed thereto;
- (b) treating the biological material with the solution obtained from step (a) at a rate and duration sufficient to prevent cells in said biological material from returning to isotonic volume prior to vitrification; and
(c) then subjecting the biological material to a pressure sufficient for vitrification upon cooling without substantial nucleation or ice crystal growth and without significant injury to the biological material. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
- (b) treating the biological material with the solution obtained from step (a) at a rate and duration sufficient to prevent cells in said biological material from returning to isotonic volume prior to vitrification; and
Specification