Method of differentiating umbilical cord tissue into a chondrogenic phenotype
View Patent ↗Cells derived from postpartum tissue and methods for their isolation and induction to differentiate to cells of a chondrogenic or osteogenic phenotype are provided by the invention. The invention further provides cultures and compositions of the postpartum-derived cells and products such as lysates related thereto. The postpartum-derived cells of the invention and products related thereto have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications, for example, in the treatment of bone and cartilage conditions such as osteoarthritis.
1. An in vitro method of inducing differentiation of a population of cells to a chondrogenic phenotype comprising:
i) obtaining an isolated homogeneous population of umbilical cord tissue-derived cells,
wherein said isolated cell population is obtained from human umbilical cord tissue substantially free of blood, can undergo at least 40 doublings in culture, maintains a normal karyotype upon passaging, expresses each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2, and HLA-A, B, C; and does not express any of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, or HLA-DR, DP, DQ, exhibits increased expression of a gene encoding interleukin 8 relative to expression of an endogenous gene encoding interleukin 8 by a human cell that is a fibroblast, a mesenchymal stem cell, or an iliac crest bone marrow cell, and exhibits increased expression of a gene encoding reticulon 1 relative to expression of an endogenous gene encoding reticulon 1 by a human cell that is a fibroblast, a mesenchymal stem cell, or an iliac crest bone marrow cell;
ii) culturing said isolated cell population in chondrogenic induction medium, wherein the induction medium comprises modified essential medium comprising L-glutamine, sodium pyruvate, L-proline, L-ascorbic acid, insulin, transferrin, and selenium; and
iii) exposing the isolated homogeneous population of cells in culture to one or more chondrogenic differentiation-inducing agents, wherein said exposing induces differentiation of said isolated cell population to a chondrogenic phenotype.
2. The method of claim 1 wherein said chondrogenic differentiation-inducing agent is transforming growth factor-beta3 (TGFbeta3).
3. The method of claim 1 , wherein said isolated cell population secretes each of the factors MCP-1, MIP1b, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, RANTES and TIMP1.
4. The method of claim 1 , wherein said isolated cell population does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb, MIP1a and VEGF.
5. The method of claim 1 wherein said isolated cell population is obtained by contacting said umbilical cord tissue with an enzyme solution of a matrix metalloprotease, a neutral protease, and a mucolytic enzyme that digests hyaluronic acid.
6. The method of claim 1 , wherein the induction medium further comprises at least one of collagen and sodium hydroxide.