Methods for Preparing and Storing Immortalized Cytolytic Natural Killer Cells
AssigneeImmunityBio, Inc.
InventorsShannyn Bessette, Syed Raza Ali, Manju Saxena
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Abstract
Provided herein are methods of preparing and storing NK-92® cells. The methods comprise storing and freezing NK-92® cells in freezing medium comprising a first composition and a second composition. Also disclosed are methods of harvesting NK-92® cells comprising collecting the cells from a cell culture and washing the collected cells in a buffer comprising albumin.
Claims26
A method of freezing cells from an immortalized non-Hodgkin's lymphoma derived cytolytic cancer cell line (CRL-2407), the method comprising:
- contacting the cells with a cell freezing medium, wherein the cell freezing medium comprises a first composition and a second composition, wherein the first composition comprises:
- (a) one or more electrolytes selected from the group consisting of potassium ions, sodium ions, magnesium ions, and calcium ions;
- (b) one or more macromolecular agents selected from the group consisting of human albumin, polysaccharide and colloidal starch;
- (c) a pH buffer;
- (d) at least one sugar;
- (e) at least one sugar alcohol;
- (f) an impermeant agent selected from the group consisting of lactobionate, gluconate, citrate and glycerophosphate;
- and
- (g) DMSO;
- and wherein the second composition comprises albumin.
2-27. (canceled)
A cell culture of cells from an immortalized non-Hodgkin's lymphoma derived cytolytic cancer cell line (CRL-2407) comprising:
- the cells and a cell freezing medium, wherein the cell freezing medium comprises a first composition and a second composition, wherein the first composition comprises:
- (a) one or more electrolytes selected from the group consisting of potassium ions, sodium ions, magnesium ions, and calcium ions;
- (b) one or more macromolecular agents selected from the group consisting of human albumin, polysaccharide and colloidal starch;
- (c) a pH buffer;
- (d) at least one sugar;
- (e) at least one sugar alcohol;
- (f) an impermeant agent being at least one member selected from the group consisting of lactobionate, gluconate, citrate and glycerophosphate;
- and
- (g) DMSO;
- and
- wherein the second composition comprises albumin.
29-44. (canceled)
A cell freezing medium produced by mixing a first composition and a second composition at a one to one ratio, wherein the first composition comprises:
- (a) one or more electrolytes selected from the group consisting of potassium ions, sodium ions, magnesium ions, and calcium ions;
- (b) one or more macromolecular agents selected from the group consisting of human albumin, polysaccharide and colloidal starch;
- (c) a pH buffer;
- (d) at least one sugar;
- (e) at least one sugar alcohol;
- (f) an impermeant agent selected from the group consisting of lactobionate, gluconate, citrate and glycerophosphate;
- and
- (g) DMSO;
- and wherein the second composition comprises albumin.
A method of harvesting cells from an immortalized non-Hodgkin's lymphoma derived cytolytic cancer cell line (CRL-2407) comprising collecting the cells from a cell culture and washing the collected cells by resuspending the cells in a buffer comprising 1-5% albumin.
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Dependencies are inferred from the claim text.