Compositions and methods for treating and repairing tendons

Abstract

The present invention relates to compositions and methods utilizing hair follicle derived Non-Bulbar Dermal Sheath cells for use in the treatment or prevention of the tendon injuries.

1 . A method for isolating non-bulbar dermal sheath (NBDS) cells, comprising:
 (a) preparing vital hair;   (b) cleaving the hair prepared in step (a) to remove the hair follicle bulb and obtain an isolated dermal sheath;   (c) isolating NBDS tissue from the dermal sheath prepared in step (b); and   (d) cultivating the isolated NBDS tissue of step (c) to produce the isolated NBDS cells.   
     
     
         2 . The method according to  claim 1  wherein said vital hair is obtained by biopsy from the occipital scalp of a subject. 
     
     
         3 . The method according to  claim 1  wherein said hair is cleaved utilizing a micromanipulator and scalpel. 
     
     
         4 . The method according to  claim 1 , further comprising the step of conducting enzymatic digestion of said isolated Non-Bulbar Dermal Sheath tissue. 
     
     
         5 . The method according to  claim 4  wherein said enzymatic digestion is conducted with collagenase. 
     
     
         6 . The method according to  claim 1  wherein said NBDS cells are cultivated over multiple passages in either serum containing or serum-free media. 
     
     
         7 - 21 . (canceled) 
     
     
         22 . The method according to  claim 6  wherein said NBDS cells are expanded in culture for 1, 2, 3, 4, 5, 10, 20 or more passages. 
     
     
         23 . The method according to  claim 6  wherein said NBDS cells are cultured over at least 1, 2, 3, 4, 5, 6, 10, or 20 passages. 
     
     
         24 . The method according to  claim 23  wherein said NBDS cells are placed into dishes or flasks which allow the NBDS cells to adhere, and with each passage non-adherent cells are removed, and the remaining adherent cells are released, followed by addition of fresh media. 
     
     
         25 . The method according to  claim 24  wherein the adherent cells are further isolated by flow cytometry. 
     
     
         26 . The method according to  claim 6  wherein said NBDS cells are passaged for at least 2, 3, 4, 5, 10 or more passages, the cultured cells are analysed in order to ascertain whether there is a sufficient quantity of NBDS cells, and whether the cells have been sufficiently isolated from contaminating cells. 
     
     
         27 . The method according to  claim 6  wherein said NBDS cells are passaged for a number of passages until approximately 5 to 100 million cells are obtained. 
     
     
         28 . The method according to  claim 27 , wherein the obtained cells are washed several times, trypsinized, and resuspended in cell transportation medium (CTM), and the resuspended cells are counted and adjusted to provide a final concentration of 20 million cells/ml and then stored in liquid nitrogen. 
     
     
         29 . The method according to  claim 6  wherein said NBDS cells are passaged for four passages to obtain approximately 100 million cells. 
     
     
         30 . A composition comprising the isolated NBDS cells prepared by the method of  claim 1 . 
     
     
         31 . A composition comprising the isolated NBDS cells prepared by the method of  claim 6 . 
     
     
         32 . A composition comprising the isolated NBDS cells prepared by the method of  claim 27 . 
     
     
         33 . A composition comprising the isolated NBDS cells prepared by the method of  claim 29 .