Methods and compositions for in vitro and in vivo chondrogenesis
Aspects of the present invention include methods and compositions related to the production, identification and use of embryonic progenitor cell lines that are capable of undergoing chondrogenesis. A number of exemplary chondrogenic cell lines derived from primordial stem cells are disclosed. The chondrogenic cell lines described herein are robust, can expand for >40 passages, and have site-specific purity, thus providing for compositions and methods of producing diverse cartilage types with unique molecular compositions for use in research and therapy.
1. A method of producing cartilage, the method comprising culturing a clonally-purified embryonic progenitor cell line; wherein said cell line is negative for the expression of one or more of the genes selected from the group consisting of: CD74, CD90, CD166, ITGA2, and KCNK2; wherein said cell line is selected from the group consisting of: SM30, E15, 4D20.8, 7SM0032, MEL2, SK11 and combinations thereof; and wherein said culturing comprises culture under cartilage producing conditions for an amount of time effective to induce cartilage production from said cells.
2. The method of claim 1 , wherein the embryonic progenitor cell line is negative for the expression of CD74.
3. The method of claim 2 , wherein the embryonic progenitor cell line is further negative for the expression of one or more genes selected from the group consisting of: HOX genes and PITX1.
4. The method of claim 3 , wherein the embryonic progenitor cell line generates cartilage in the absence of COL10A1 marker expression.
5. The method of claim 1 , wherein the cartilage producing condition is selected from one or more of the group consisting of: chondrocyte culture conditions; impregnating the embryonic progenitor cell line into synthetic matrices or biological resorbable immobilization vehicles; and placing the embryonic progenitor cell line into a molded structure.
6. The method of claim 1 , wherein the method further comprises transplanting the cartilage produced by the embryonic progenitor cell line to a subject.