Controlled reversible poration for preservation of biological materials
AssigneeThe General Hospital Corporation
InventorsMehmet Toner, Michael Russo, Robert Bieganski
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Abstract
A preservation method for biological material having cell membranes includes reversibly porating the cell membranes; loading a bio-protective agent having bio-preservation properties to a predetermined intracellular concentration; preparing the bio-protective agent loaded biological material for storage; storing the biological material; recovering the stored biological material from storage; and reversing the cell membrane poration. H5 α-toxin, a genetically engineered mutant of Staphylococcus aureus α-hemolysin, may be used as a porating agent. Non-permeating sugars such as trehalose and sucrose may be used as the bio-protective agent.
Claims30
A method for preserving nucleated cells having lipid membranes, comprising:
- a. Reversibly porating the lipid membranes of the nucleated cells;
- b. Loading the porated nucleated cells with a bio-preserving agent having bio-preservation properties to a predetermined intracellular concentration to preserve a cellular material, the predetermined intracellular concentration of the bio-preserving agent being less than or equal to about 1.0 M;
- c. Preparing the bio-preservation agent loaded nucleated cells for storage by a method selected from the group consisting of cryopreserving, freeze drying, and drying without the use of a freezing step;
- and
- d. Storing the prepared nucleated cells so that they can be recovered to a viable state in which the nucleated cells survive and grow.
A method for preserving mammalian cells having lipid membranes, comprising:
- a. Applying a membrane toxin to reversibly porate the lipid membranes of the mammalian cells;
- b. Loading the porated mammalian cells with an agent having bio-preservation properties to a predetermined intracellular concentration sufficient for preserving the cellular material, the agent comprising a non-permeating sugar and the predetermined intracellular concentration of the agent being less than or equal to about 1.0 M;
- c. Preparing the bio-preservation agent loaded mammalian cells for storage by a method selected from the group consisting of cryopreserving, freeze drying, and drying without the use of a freezing step;
- and
- d. Storing the prepared mammalian cells so that they can be recovered to a viable state in which the mammalian cells survive and grow.
Dependencies are inferred from the claim text.