Derivation of fibrochondrocytes from progenitor cells
View Patent ↗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 method of treating a subject having a meniscus tissue defect, the method comprising:
(a) implanting into a subject in need thereof, at a site of a meniscus tissue defect, a scaffold comprising
(i) an anatomically correct model of a meniscus,
(ii) connective tissue growth factor (CTGF) encapsulated in a first microsphere in an outer region of the scaffold, and
(iii) transforming growth factor β3 (TGFβ3) encapsulated in a second microsphere in an inner region of the scaffold;
(b) inducing migration of a progenitor cell into or onto the scaffold;
(c) forming chondrocytes or chondrocyte-like cells in the inner region of the meniscus shaped scaffold; and
(d) forming fibroblasts or fibroblast-like cells in the outer region of the meniscus shaped scaffold,
wherein
the first microsphere releases CTGF before the second microsphere releases TGFβ3, and
the scaffold does not comprise a transplanted cell prior to implant.
2. The method of claim 1 , wherein the scaffold comprises a biocompatible matrix material.
3. The method of claim 1 , wherein the scaffold comprises poly(lactic-co-glycolic acid) (PLGA) or polycaprolactone (PCL).
4. The method of claim 1 , wherein the scaffold comprises at least one physical channel.
5. The method of claim 1 , wherein the scaffold is an anatomically correct model of a knee meniscus.
6. The method of claim 1 , wherein the scaffold further comprises:
a fibroblastic induction supplement comprising ascorbic acid; or
a chondrogenic induction supplement comprising one or more of ITS+1 solution, sodium pyruvate, ascorbic acid 2-phosphate, proline, or dexamethasone.
7. The method of claim 1 , wherein 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.
8. The method of claim 1 , wherein CTGF has a concentration of about 100 ng/mL; and TGFβ3 has a concentration of about 10 ng/mL.
9. The method of claim 1 , wherein the first microsphere is a 50:50 poly(lactic-co-glycolic acid) (PLGA) microsphere and the second microsphere is a 75:25 PLGA microsphere.
10. The method of claim 1 , wherein the number of formed fibrochondrocytes or fibrochondrocyte-like cells in the scaffold is at least about 100% greater than the number of fibrochondrocytes or fibrochondrocyte-like cells formed under conditions not comprising CTGF and TGFβ3.
11. The method of claim 1 , wherein the fibrochondrocytes or fibrochondrocyte-like cells display one or more of: increased collagen (COL) deposition; increased proteoglycan (PG) deposition; increased glycosaminoglycan (GAG) deposition; increased proCOL-I+; or increased proCOL-IIα+, as compared to the progenitor cell.
12. The method of claim 1 , wherein the progenitor cell is a mesenchymal stem cell.
13. The method of claim 1 , wherein the progenitor cell is a human mesenchymal stem cell.
14. The method of claim 1 , wherein the first microsphere releases CTGF faster than the second microsphere releases TGFβ3.