Abstract: The invention discloses a multi-layer cell freezing vial and method of obtention thereof. The method of obtention is a cell freezing and thawing process that avoids the need of incorporating new fresh medium to the thawed cells for the dilution of the cryoprotective agent. This is achieved by the previous freezing of an extra layer of culture medium in addition to the frozen cell solution containing said cryoprotective agent. At the time of thawing, the culture medium and the cell solution mix themselves, turning the concentration of said cryoprotective agent to a non-toxic dilution and achieving the effective exclusion of said cryoprotective agent from the living cells.

Abstract: The present invention is a method of freezing cells comprising the steps of incubating said cells in a solution comprising a cryoprotective agent, concentrating the cells resulting from the previous step withdrawing the eluent essentially free of cells, and freezing the resulting concentrated cells. The cells frozen by the invention's method render a high post-thawing viability, reduce cryoprotectant related toxic events and promote cells life in a suspension state after thawing. The invention also comprises the container comprising the frozen cells.