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

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.

1 . A method of treating an injury or disease in a mammal, comprising providing to the mammal a cell population comprising adipose-tissue derived stem cells, wherein said cell population was prepared by a cell processing 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 cell processing method does not include isolating stem cells separated in (b) from other cells separated in (b).   
     
     
         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 the adipose tissue-derived stem cells are autologous to the mammal being treated. 
     
     
         7 . 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. 
     
     
         8 . The method of  claim 1 , wherein said cell population further comprises one or more other tissue components. 
     
     
         9 . The method of  claim 8 , wherein the one or more other tissue components comprise: extracellular matrix polypeptides, proteoglycans, lipids, cytokines, or growth factors. 
     
     
         10 . The method of  claim 1 , wherein the disease or disorder is selected from the group consisting of: inflammatory diseases or disorders, cardiovascular diseases, nervous system diseases, tumors, demyelinating diseases, digestive system diseases, endocrine system diseases, reproductive system diseases, hemic and lymphatic diseases, immunological diseases, mental disorders, musculoskeletal diseases, neuromuscular diseases, metabolic diseases, skin and connective tissue diseases, and urological diseases. 
     
     
         11 . The method of  claim 1 , wherein the injury is a wound selected from the group consisting of: abrasions, avulsions, blowing wounds, incised wounds, burns, contusions, puncture wounds, surgical wounds and subcutaneous wounds. 
     
     
         12 . A method of treating a disease or disorder in a mammal, comprising:
 (a) preparing a cell population comprising adipose tissue-derived stem cells for introducing into a mammal by a method comprising:
 (i) processing adipose tissue obtained from a mammal to release cells therein, wherein the adipose tissue comprises stem cells, wherein said processing releases cells and a fat layer; and 
 (ii) separating the cells released in (i) from the fat layer, 
   wherein the preparing does not include isolating stem cells separated in (a)(ii) from other cells separated in (a)(ii), thereby preparing the cell population comprising adipose tissue-derived stem cells; and   (b) providing the cell population of (a)(ii) to the mammal.   
     
     
         13 . The method of  claim 12 , wherein the processing 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.   
     
     
         14 . The method of  claim 12 , wherein the wherein the separating comprises one or more procedures selected from the group consisting of:
 use of a density gradient;   centrifugation; and   filtration.   
     
     
         15 . The method of  claim 12 , wherein the adipose tissue-derived stem cells are autologous to the mammal being treated. 
     
     
         16 . The method of  claim 12 , wherein the 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. 
     
     
         17 . The method of  claim 12 , wherein the cell population further comprises one or more other tissue components. 
     
     
         18 . The method of  claim 17 , wherein the one or more other tissue components comprise: extracellular matrix polypeptides, proteoglycans, lipids, cytokines, or growth factors. 
     
     
         19 . The method of  claim 12 , further comprising:
 (c) freezing the cell population in a physiologically compatible buffer before step (b).   
     
     
         20 . The method of  claim 12 , further comprising:
 (c) suspending the cell population in a physiologically compatible buffer before step (b).   
     
     
         21 . The method of  claim 12 , wherein the disease or disorder is selected from the group consisting of: inflammatory diseases or disorders, cardiovascular diseases, nervous system diseases, tumors, demyelinating diseases, digestive system diseases, endocrine system diseases, reproductive system diseases, hemic and lymphatic diseases, immunological diseases, mental disorders, musculoskeletal diseases, neuromuscular diseases, metabolic diseases, skin and connective tissue diseases, and urological diseases. 
     
     
         22 . The method of  claim 12 , wherein the injury is a wound selected from the group consisting of: abrasions, avulsions, blowing wounds, incised wounds, burns, contusions, puncture wounds, surgical wounds and subcutaneous wounds. 
     
     
         23 . The method of  claim 12 , wherein the disease or disorder is a musculoskeletal injury or disease. 
     
     
         24 . The method of  claim 12 , wherein the musculoskeletal injury or disease is a sprain, strain, dislocation, bruising, tear, microtear, contusion, bursitis, tendonitis, articular cartilage injury or fracture in a tendon, ligament, cartilage, bone, or hoof laminae.