Methods of preparing and using novel stem cell compositions and kits comprising the same
The present invention provides novel stem cell compositions having significant therapeutic and practical advantages, as well as methods of preparing and using such compositions for the treatment and prevention of injury and disease in patients. The invention may be applied to stem cell populations isolated from a wide variety of animals, including humans, and tissues. In particular applications, the invention is used to prepare a stem cell composition from a collagen-based tissue, such as adipose tissue, isolated from a patient, and the stem cell composition is subsequently provided to a site of actual or potential injury in the patient. The invention further includes related kits comprising the stem cell compositions, which are remarkably stable and retain viability and efficacy during storage and shipment.
1. A method of preparing a cell population comprising adipose-tissue derived stem cells useful for the treatment of an injury or disease, said method comprising:
(a) processing adipose tissue obtained from a mammal to release cells therein, wherein said processing releases cells and a fat layer; and
(b) separating cells released in (a) from the fat layer, thereby preparing the cell population comprising adipose tissue-derived stem cells;
wherein said method does not include isolating stem cells separated in (b) from other cells separated in (b), and
wherein the cell population obtained from (b) is useful for the treatment of an injury or disease and shows less than 20% reduction in viability following storage under refrigeration at 4° C. for 20-24 hours.
2. The method of claim 1 , wherein the processing of (a) comprises one or more procedures selected from the group consisting of: physical cutting or mincing of the adipose tissue; treating the adipose tissue with an enzyme that facilitates the release of cells; exposing the adipose tissue to ultrasonic energy; and treating the adipose tissue with perfluorocarbons.
3. The method of claim 1 , wherein the processing of (a) comprises physical cutting or mincing of the adipose tissue.
4. The method of claim 1 , wherein the separating of (b) comprises one or more procedures selected from the group consisting of: use of a density gradient; centrifugation; and filtration.
5. The method of claim 1 , wherein:
the processing of (a) comprises one or more procedures selected from the group consisting of: physical cutting or mincing of the adipose tissue; treating the adipose tissue with an enzyme that facilitates the release of cells; exposing the adipose tissue to ultrasonic energy; and treating the adipose tissue with perfluorocarbons; and
the separating of (b) comprises one or more procedures selected from the group consisting of: use of a density gradient; centrifugation; and filtration.
6. The method of claim 1 , wherein said cell population further comprises one or more cells selected from the group consisting of: red blood cells, white blood cells, fibroblasts, neutrophils, monocyte/macrophages, and basophils.
7. The method of claim 1 , wherein said cell population further comprises one or more other tissue components.
8. The method of claim 7 , wherein the one or more other tissue components comprise: extracellular matrix polypeptides, proteoglycans, lipids, cytokines, or growth factors.
9. The method of claim 1 , further comprising:
(c) freezing the cell population.
10. The method of claim 1 , further comprising:
(c) suspending the cell population in a physiologically compatible buffer.
11. The method of claim 1 , further comprising:
(c) placing the cell population in a container.
12. The method of claim 11 , wherein the container is a syringe.
13. The method of claim 11 , further comprising shipping the container comprising the cell population to a physician or veterinarian.
14. The method of claim 1 , further comprising storing the cell population under refrigeration.