Methods of preparing and using novel stem cell compositions and kits comprising the same

• US 10,668,105 B2
• Filed: 08/19/2016
• Issued: 06/02/2020
• Est. Priority Date: 10/08/2003
• Status: Active Grant

• Current Assignee: Vetstem Incorporated
• Original Assignee: Vetstem Biopharma, Inc.
• Inventors: Harman, Jr., Robert J., Sand, Theodore T.
• Primary Examiner(s): Singh, Satyendra K
• Application Number:
  US15/241,656
• Publication Number:
  US 20170196912A1
• Time in Patent Office: 1,383 Days
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2509/10 Mechanical dissociation
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

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.

Claims

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.