CULTURE OF HUMAN MESENCHYMAL STEM CELLS FROM BONE MARROW AND THEIR DIFFERENTIATION INTO CARTILAGE / 解放军医学杂志

ABSTRACT

To develop the methodology to purify and culture expand mesenchymal stem cells (MSC) from human bone marrow and investigate the optimal system for MSC differentiation into chondrocytes mononuclear cells were harvested by gradient centrifugation on Percoll at density of 1 073g/ml and seeded in low glucose DMEM containing 10% fetal calf serum. The purity of cells was evaluated by flow cytometric technique. MSC of 2 passages thereafter were absorbed into a biomaterial of gelatin sponge and induced to differentiate in to chondrocytes for one week under the influence of TGF ? and other inductive agents. The feature of chondrocytes in engineered tissues was identified by toluidine blue staining at various time points. The results showed that human mesenchymal stem cells culture expanded were positive for CD166, CD29, and CD44, but negative for CD34, CD45, and HLA DR. Furthermore, when treated cells absorbed into the biomaterial were implanted subcutaneously into BALB c nude mice, they formed cartilage like tissues after 4 weeks.Our conclusions is that cultured marrow MSC have unique biological features and the capacity to differentiate into chondrocytes in vivo , so they are useful for cartilage engineering by serving as the seed cells.