Methods and compositions for the preservation of cells, tissues or organs in the vitreous state

Patent No. 6,921,633

Issued: July 26, 2005
Filed: November 18, 2002

Inventors: Baust; John M. (Candor, NY); Baust; John G. (Candor, NY); VanBuskirk; Robert (Apalachian, NY); Mathew; Aby J. (Binghamton, NY)

Assignee: Biolife Solutions Incorporated (Binghamton, NY)

The invention encompasses a method of preserving a eukaryotic cell, tissue or organ comprising: a) contacting the cell, tissue or organ with a hypothermic storage solution, wherein the solution comprises: i) a composition that inhibits apoptosis; and ii) a concentration of a vitrification composition that is sufficient for vitrification of the solution; and b) vitrifying the cell, tissue or organ, wherein the vitrification occurs both within the cell, tissue or organ and in the hypothermic storage solution comprising and comprised by the cell, tissue or organ.

The vitrification is accomplished through use of a hypothermic storage solution comprising an agent that prevents ice nucleation within the extracellular and intracellular environment thereby preventing ice formation and that has a glass transition temperature (Tg) lower than the homogeneous nucleation temperature of the solution. Reduction of the temperature of a sample in a hypothermic storage solution to below the glass transition temperature results in vitrification of the solution and the cell, tissue or organ in that solution. Under these circumstances, there is no crystalline ice formation in or around the cells as the sample becomes a solid. The inclusion of one or more anti-apoptotic agents aids in preventing the apoptotic cell death that normally occurs following this type of preservation.

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