Derivation of fibrochondrocytes from progenitor cells

Abstract

Provided herein are compositions and methods for forming fibrochondrocytes or fibrochondrocyte-like cells from progenitor cells, such as mesenchymal stem cells. One aspect provides a fibrochondrocyte culture medium including CTGF and TGFβ3, optionally encapsulated by microspheres having different release profiles. Another aspect provides a method for forming fibrochondrocytes or fibrochondrocyte-like cells from progenitor cells by culturing with CTGF and TGFβ3.

1 . A culture medium for generating fibrochondrocyte or fibrochondrocyte-like cells comprising:
 CTGF encapsulated in a first microsphere; and   TGFβ3 encapsulated in a second microsphere,   wherein the first microsphere and the second microsphere have different release profiles.   
     
     
         2 . The culture medium of  claim 1 , wherein the first microsphere is a 50:50 PLGA microsphere and the second microsphere is a 75:25 PLGA microsphere. 
     
     
         3 . The culture medium of  claim 1 , wherein:
 (i) CTGF has a concentration of about 10 to about 1000 ng/mL; and TGFβ3 has a concentration of about 1 to about 1000 ng/mL; or   (ii) CTGF has a concentration of about 100 ng/mL; and TGFβ3 has a concentration of about 10 ng/mL.   
     
     
         4 . The culture medium of  claim 1 , further comprising a fibroblastic induction supplement or a chondrogenic induction supplement. 
     
     
         5 . A method of forming fibrochondrocytes or fibrochondrocyte-like cells comprising:
 (i) contacting a progenitor cell and the culture medium of  claim 1 ; or contacting a progenitor cell with CTGF and TGFβ3 sequentially or concurrently; and   (ii) culturing the progenitor cell so as to form a fibrochondrocyte or fibrochondrocyte-like cell.   
     
     
         6 . The method of  claim 5 , wherein:
 (i) the first microsphere releases CTGF earlier or faster than the second microsphere releases TGFβ3;   (ii) the first microsphere releases CTGF later or slower than the second microsphere releases TGFβ3;   (iii) the first microsphere releases CTGF about the same time as the second microsphere releases TGFβ3;   (iv) the progenitor cell is contacted with CTGF followed sequentially by contact with TGFβ3;   (v) the progenitor cell is contacted with TGFβ3 followed sequentially by contact with CTGF; or   (vi) the progenitor cell is contacted concurrently with TGFβ3 and CTGF.   
     
     
         7 . The method of  claim 5 , wherein the culturing the progenitor cell occurs for (i) about two days to about five days; or (ii) at least about two days, at least about three days, at least about four days, or at least about five days. 
     
     
         8 . The method of  claim 5 , wherein the number of formed fibrochondrocytes or fibrochondrocyte-like cells is at least about 100% greater than the number of fibrochondrocytes or fibrochondrocyte-like cells formed under culture conditions not comprising CTGF and TGFβ3. 
     
     
         9 . The method of  claim 5 , wherein the fibrochondrocytes or fibrochondrocyte-like cells display one or more of: increased COL deposition; increased PG deposition; increased GAG deposition; increased proCOL-I+; or increased proCOL-IIα+, as compared to the progenitor cell. 
     
     
         10 . The method of  claim 5 , wherein the progenitor cell is a mesenchymal stem cell. 
     
     
         11 . The method of  claim 5 , wherein the progenitor cell is a human mesenchymal stem cell. 
     
     
         12 . A method of forming a fibrocartilage tissue comprising:
 providing a scaffold comprising an effective amount of the culture medium of  claim 1 ;   placing the scaffold in fluid communication with a progenitor cell;   inducing migration of the progenitor cell into or onto the scaffold; and   inducing formation of a fibrochondrocyte or fibrochondrocyte-like cell from the progenitor cell,   wherein, the scaffold does not comprise a transplanted cell.   
     
     
         13 . A method of treating a subject having a fibrocartilage tissue defect, the method comprising:
 implanting into a subject in need thereof a scaffold comprising an effective amount of the culture medium of  claim 1 ;   wherein
 the scaffold does not comprise a transplanted cell prior to implant; 
 the effective amount of the culture medium induces migration of a progenitor cell into or onto the scaffold; and 
 the effective amount of the culture medium induces formation of a fibrochondrocyte or fibrochondrocyte-like cell from the progenitor cell. 
   
     
     
         14 . A fibrocartilage tissue construct comprising:
 a scaffold comprising an effective amount of the culture medium of  claim 1 ,   wherein,
 the scaffold does not comprise a transplanted cell; 
 the effective amount of the culture medium can induce migration of a progenitor cell into or onto the scaffold when the scaffold is in fluid communication with the progenitor cell; and 
 the effective amount of the culture medium can induce formation of an fibrochondrocyte or fibrochondrocyte-like cell from the progenitor cell. 
   
     
     
         15 . The method of  claim 13 , wherein the scaffold comprises a biocompatible matrix material. 
     
     
         16 . The method of  claim 13 , wherein the scaffold comprises poly(lactic-co-glycolic acid) (PLGA). 
     
     
         17 . The method of  claim 13 , wherein the scaffold comprises at least one physical channel. 
     
     
         18 . The method of  claim 13 , wherein the scaffold comprises at least two layers. 
     
     
         19 . The method of  claim 16 , wherein the scaffold comprises a first layer and a second layer, the first layer comprises CTGF and the second layer comprises TGFβ3.