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 preparing a purified cell population comprising stem cells for introducing into a patient, comprising:
 (a) obtaining adipose tissue from said patient;   (b) processing said adipose tissue to separate cells therein from other tissue components; and   (c) purifying the separated cells from other tissue components;   wherein said method does not include isolating stem cells from other purified cells, thereby preparing a purified cell population comprising adipose tissue-derived stem cells.   
     
     
         2 . The method of  claim 1 , wherein said purified cell population further comprises one or more cells selected from the group consisting of: red blood cells, white blood cells, fibroblasts, fibroblast-like cells, neutrophils, monocyte/macrophages, and basophils. 
     
     
         3 . The method of  claim 1 , wherein said purified cell population further comprises one or more tissue components selected from the group consisting of: extracellular matrix polypeptides or fragments thereof, proteoglycans, cytokines, and growth factors. 
     
     
         4 . The method of  claim 3 , wherein said extracellular matrix polypeptide is selected from the group consisting of: collagen, thrombospondin, fibronectin, vitronectin, cytotactin, laminin and integrins. 
     
     
         5 . The method of  claim 1 , further comprising suspending said purified cells in a physiologically compatible buffer. 
     
     
         6 . The method of  claim 5 , further comprising placing said purified cells in a syringe. 
     
     
         7 . The method of  claim 1 , further comprising freezing said isolated cells in freezing medium. 
     
     
         8 . The method of  claim 1 , wherein said processing comprises one or more procedures selected from the group consisting of:
 (a) mincing the adipose tissue;   (b) treating the adipose tissue with an enzyme that facilitates the release of cells from other tissue components;   (c) exposing the adipose tissue to ultrasonic energy; and   (d) treating the adipose tissue with perflurocarbons.   
     
     
         9 .- 10 . (canceled) 
     
     
         11 . The method of  claim 1 , wherein said patient is a human. 
     
     
         12 . The method of  claim 1 , wherein said patient is a non-human animal. 
     
     
         13 .- 19 . (canceled) 
     
     
         20 . The method of  claim 12 , wherein said adipose tissue is obtained from the tail head region of the animal. 
     
     
         21 . A method of preparing a composition comprising purified adipose tissue-derived stem cells for introducing into an animal, comprising:
 (a) obtaining adipose tissue from the tail head region of said animal;   (b) processing said adipose tissue to separate cells therein from other tissue components; and   (c) purifying the separated cells from other tissue components,   thereby preparing a composition comprising purified adipose tissue-derived stem cells.   
     
     
         22 . The method of  claim 21 , wherein said composition further comprises one or more cells selected from the group consisting of: red blood cells, white blood cells, fibroblasts, fibroblast-like cells, neutrophils, monocyte/macrophages, and basophils. 
     
     
         23 . The method of  claim 21 , wherein said composition further comprises one or more tissue components selected from the group consisting of: extracellular matrix polypeptides or fragments thereof, proteoglycans, cytokines, and growth factors. 
     
     
         24 . The method of  claim 23 , wherein said extracellular matrix polypeptide is selected from the group consisting of: collagen, thrombospondin, fibronectin, vitronectin, cytotactin, laminin and integrins. 
     
     
         25 .- 28 . (canceled) 
     
     
         29 . The method of  claim 21 , wherein said processing comprises one or more procedures selected from the group consisting of:
 (a) mincing the adipose tissue;   (b) treating the adipose tissue with an enzyme that facilitates the release of cells from other tissue components;   (c) exposing the adipose tissue to ultrasonic energy; and   (d) treating the adipose tissue with perflurocarbons.   
     
     
         30 .- 31 . (canceled) 
     
     
         32 . A method of providing a composition comprising purified stem cells for introducing into a patient, comprising:
 (a) processing collagen-based tissue obtained from a patient to separate cells therein from other tissue components;   (b) purifying the separated cells; and   (c) placing said separated cells in a container, thereby providing a composition comprising purified stem cells.   
     
     
         33 .- 150 . (canceled)