Methods for the cryopreservation of cells

摘要

The present invention features novel methods for the cryopreservation of mammalian cell that combine the advantages of the slow-freezing and vitrification approaches while avoiding their shortcomings. Generally, the methods include the use of a capillary tube made of a thermally conductive wall material and a thin wall such that the ratio of the thermal conductivity of the wall material to the wall thickness is at least 1,000-500,000. The solution is then exposed to temperatures equal to or less than −80° C. and the vitrification solution containing the mammalian cells is cooled at a rate equal to or greater than 30,000-100,000,000° C./minute. The exposure of the capillary tube with a thermally conductive and thin wall allows for vitrification of the solution in the absence of ice formation. Cryoprotectants can also be added to the vitrification solution to further prevent ice formation.

主权项

1. A method for cryopreserving a mammalian cell, the method comprising:
(a) suspending said cell in a vitrification solution comprising one or more cryoprotective agents, wherein a concentration of the combination of all cryoprotective agents in said vitrification solution is less than or equal to 4 M; (b) placing said cell in said vitrification solution in a microcapillary tube, wherein said microcapillary tube has a wall made of a material having a thermal conductivity of at least 0.2 W m−1K−1; and (c) cooling said cell in said vitrification solution in said microcapillary tube to a temperature less than or equal to a glass transition temperature of said vitrification solution at a rate greater than 200,000° C./min, wherein said cooling causes vitrification of said cell.