Method of freezing cells
AssigneeZf Biotox, S.L.
InventorsRoberto Hernan Izquierdo, Natalia Gallot Escobal, Antonio Cruz Pacheco
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Abstract
The invention discloses a multi-layer cell freezing vial and method of ohtention 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 ceil 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.
Claims23
Method of freezing cells or cell-like materials or non-human embryos, said method comprising the steps of:
- a) freezing a layer of biologically acceptable liquid diluent in a container;
- b) adding onto said frozen diluent a layer of said cells or cell-like materials or non-human embryos in form of a solution, the medium of said solution comprising at least one cryoprotective agent in suitable concentration;
- and
- c) freezing the result of the previous step to obtain a double-layer frozen body with the frozen cells or cell-like materials or non-human embryos solution on top;
- or
- a) freezing a layer of cells or cell-like materials or non-human embryos in form of a solution, the medium of said solution comprising at least one cryoprotective agent in suitable concentration, in a container;
- b) adding onto said frozen solution a layer of biologically acceptable liquid diluent;
- and
- c) freezing the result of the previous step to obtain a double-layer frozen body with the frozen diluent on top.
Dependencies are inferred from the claim text.