Abstract: Novel methods, compositions, and devices for achieving optimal cooling of living cells during cryopreservation are disclosed. In one aspect, the method comprises gradually cooling the cell to a first predetermined temperature, followed by rapidly cooling the cell to a second predetermined temperature. In another aspect, a device is described for achieving a desired cooling rate for a cell, comprising a first container for holding a cell, a second container for holding the first container, and optionally a frame for holding the first container in a spaced apart relationship with the second container. The method of the invention comprises placing cells into the first container, placing the first container in the second container and sealing the second container, and placing the second container in a suitable cooling device. In yet another aspect, novel cryoprotectant compositions are provided comprising conventional cryoprotectant plus one or more high molecular weight cryoprotectants.

Abstract: Novel methods and devices for removing cryoprotectant from cryoprotectant-containing liquids, and from cells residing therein, are disclosed. In one aspect, the method comprises passing the cryoprotectant-containing liquid through at least one semipermeable hollow fiber membrane contained in a hollow module in a first direction, while passing a liquid which is substantially free of cryoprotectant through the hollow module in a second direction to remove cryoprotectant across a diffusion gradient. In another aspect, a device is described for removing cryoprotectant from a liquid, comprising a hollow module with at least one semipermeable hollow fiber membrane therein for accomplishing such counter-current diffusion removal of cryprotectant. A software program is also provided for predicting optimal flow rates through the device of the invention, thereby allowing optimal cryoprotectant removal regardless of the cryoprotectant used or the material from which the semipermeable hollow fiber membrane is fabricated.