Detailed Study during Cartilage Formation by Human Mesenchymal Stern Cells In a Three Dimentional Culture and Implantation.

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Human mesenchymal stem cells (hMSCs) have the potential to differentiate into the lineages of mesenchymal tissues. The aim of this study Is to clarify the detailed process of differentiation In hMSCs. We examined cartilage formation of human mesenchymal stem cells using three dimentional culture and implantation.Three dimentional culture -alginate bead culture-: hMSCs were maintained in alginate beads. We examined them by light microscope, transmission electron microscope and immunocryo-ultramicrotomy. hMSCs produced collagen type II, X, proteoglycan and osteocalcin at day 4 in the case of alginate bead culture. Thereafter electron-dense particles, composed of hydroxyapatite, appeared.Three dimentional culture -pellet culture-: In the case of pellet culture, hMSCs were connected with junctional complex at day 1. At day 7, hMSCs produced collagen type I, II, X and proteoglycan. During day 14-21, the expression of collagen type II decreased.Implantation of MSO-derived cartilages into the rabbit cartilage defect.: We cultured MSCs as a micromass in chondrogenesis medium for 14 days and differentiated MSCs into cartilage in vitro. Thereafter, MSC-derived cartilages were press fitted into the rabbit cartilage defects. Eight weeks after implantation, we observed the cartilage defects were repaired.