Stem cells from urine and methods for using the same

Abstract

Provided herein are stem cells and methods for producing a culture of stem cells from urine. The stem cells may be differentiated into an osteogenic, chondrogenic, adipogenic, endothelial, neurogenic or myogenic lineage. Methods of use of the cells are provided, including methods of treating a subject in need of a cell based therapy.

1 . A method for producing a culture of differentiated cells comprising:
 providing urine stem cells isolated from a urine sample; and then   differentiating said urine stem cells, wherein said differentiating is osteogenic, chondrogenic, adipogenic, endothelial, neurogenic or myogenic;   to produce said culture of differentiated cells.   
     
     
         2 . The method of  claim 1 , wherein said differentiating is osteogenic, and wherein said culture of differentiated cells comprises mineralized bone, calcium deposition, osteocalcin, Runx2, alkaline phosphatase, or a combination thereof. 
     
     
         3 . The method of  claim 1 , wherein said differentiating is adipogenic, and wherein said culture of differentiated cells express the transcription factor PPARγ, lipoprotein lipase (LpL), or a combination thereof. 
     
     
         4 . The method of  claim 1 , wherein said differentiating is chondrogenic, and wherein said culture of differentiated cells express Sox9, collagen II, aggrecan, or a combination thereof. 
     
     
         5 . The method of  claim 1 , wherein differentiating is myogenic, and wherein said culture of differentiated cells express actin, desmin, calponin, myosin, or a combination thereof. 
     
     
         6 . The method of  claim 1 , wherein differentiating is endothelial, and wherein said culture of differentiated cells express vWF, CD 31, or a combination thereof. 
     
     
         7 . The method of  claim 1 , wherein differentiating is neurogenic, and wherein said culture of differentiated cells express nestin. 
     
     
         8 . The method of  claim 1 , wherein said urine stem cells are human or mammalian cells. 
     
     
         9 . An isolated differentiated cell produced by a method of  claim 1 . 
     
     
         10 . An isolated urine stem cell, wherein said cell is c-kit positive, and wherein said cell can differentiate into two or more lineages selected from the group consisting of: bone, cartilage, fat, endothelium, nerve and muscle. 
     
     
         11 . The cell of  claim 10 , wherein said cell is positive for a marker selected from: CD133, SSEA-A, CD90, CD73, CD105, pericyte CD146 (MCAM), NG2, PDGF-Reeeptorβ (PDGF-Rβ), and combinations thereof, and wherein said cell is negative for a marker selected from CD31, CD34, CD45, and combinations thereof. 
     
     
         12 . The cell of  claim 10 , wherein said cell expresses telomerase. 
     
     
         13 . The cell of  claim 10 , wherein said cell has undergone hypoxia preconditioning by culturing in a 1-2% O 2  environment. 
     
     
         14 . A method of providing a seeded tissue scaffold, said method comprising the steps of:
 providing the isolated urine stem cell of  claim 10 ;   differentiating said urine stem cell, wherein said differentiating is osteogenic, chondrogenic, adipogenic, endothelial, neurogenic or myogenic, to produce differentiated cells; and   seeding said differentiated cells onto a biocompatible tissue scaffold.   
     
     
         15 . The method of  claim 14 , wherein said tissue scaffold comprises a collagen matrix. 
     
     
         16 . The method of  claim 14 , wherein said tissue scaffold comprises a synthetic polymer. 
     
     
         17 . The method of  claim 14 , wherein said tissue scaffold comprises a synthetic polymer selected from the group consisting of: polyglycolic acid (PGA), polylactic acid (PLA), and polylactic-co-glycolic acid (PLGA). 
     
     
         18 . The method of  claim 14 , wherein said tissue scaffold comprises a cellulose polymer. 
     
     
         19 . A seeded tissue scaffold comprising a biocompatible tissue scaffold and urine stem cells. 
     
     
         20 . The seeded tissue scaffold of  claim 19 , wherein said biodegradable tissue scaffold comprises a collagen matrix. 
     
     
         21 . The seeded tissue scaffold of  claim 19 , wherein said biodegradable tissue scaffold comprises a synthetic polymer. 
     
     
         22 . The seeded tissue scaffold of  claim 19 , wherein said biodegradable tissue scaffold comprises a synthetic polymer selected from the group consisting of: polyglycolic acid (PGA), polylactic acid (PLA) and polylactic-co-glycolic acid (PLGA). 
     
     
         23 . The seeded tissue scaffold of  claim 19 , wherein said biodegradable tissue scaffold comprises a cellulose polymer. 
     
     
         24 . A method of treating a subject in need thereof, said method comprising the steps of:
 providing a tissue substrate, wherein said substrate comprises urine stem cells;   differentiating said urine stem cells, wherein said differentiating is osteogenic, chondrogenic, adipogenic, endothelial, neurogenic or myogenic, to produce differentiated cells;   seeding said differentiated cells onto a biocompatible substrate; and   transplanting said substrate into said patient.   
     
     
         25 . The method of  claim 24 , wherein said substrate comprises a collagen matrix. 
     
     
         26 . The method of  claim 24 , wherein said substrate comprises a synthetic polymer. 
     
     
         27 . The method of  claim 24 , wherein said substrate comprises a synthetic polymer selected from the group consisting of: polyglycolic acid (PGA), polylactic acid (PLA) and polylactic-co-glycolic acid (PLGA). 
     
     
         28 . Cultured smooth muscle tissue produced by the process of  claim 5 . 
     
     
         29 . The cultured muscle tissue of  claim 28 , wherein said tissue is characterized by a contractile response to calcium ionophore in vitro. 
     
     
         30 . The cultured muscle tissue of  claim 28 , wherein said tissue is suturable. 
     
     
         31 . The cultured muscle tissue of  claim 28 , wherein said tissue contracts upon administration of a contraction agonist four weeks after of in vivo implantation. 
     
     
         32 . A method of treating stress urinary incontinence (UI) or vesicoureteral reflux (VCR) in a subject in need thereof, comprising:
 administering urine stem cells to said subject in a treatment effective amount.   
     
     
         33 . The method of  claim 32 , wherein said administering is carried out by subcutaneous administration. 
     
     
         34 . The method of  claim 32 , wherein said urine stem cells are tranfected with a nucleic acid encoding VEFG. 
     
     
         35 . The method of  claim 32 , further comprising administering endothelial cells. 
     
     
         36 . The method of  claim 32 , wherein said cells are provided in a pharmaceutically acceptable carrier. 
     
     
         37 . The method of  claim 36 , wherein said carrier comprises a collagen gel, a hydrogel, a temperature sensitive gel or a hyaluronic acid gel. 
     
     
         38 . The method of  claim 32 , wherein said administering is carried out by endoscopic injection. 
     
     
         39 . The method of  claim 32 , wherein said subject is in need of treatment for stress urinary incontinence (SUI).