PLEGFP-N1 Modified BMSCs In The Reconst-r Uction Of Bone Defect

OBJECTIVE: To investigate the proliferative and multilineage potential cells of MSCs in vito, as seeding cells of tissue engineering. The expression of EGFP modified MSCs in vito and in the reconstruction of bone defect.METHODS: The rabbit MSCs were aspirated from the iliac crests.Mononuclear cells were separated by centrifugation in Percoll solution( 1.073g/ml) followed by the method of Pitenger M.F. and Majumda MK. Then they were seeded in DMEM-LG supplemented with 15% fetal calf-serum at a concentration of 2 × 10~5 cells/cm~2 in culture flask.In order to investigare the biological charateristics of MSCs.Amplification and recombination retroviral vector PLNCX-EGFP in order to gain albumen progression to infect incasing cells and MSC_S transfection.Induce MSC_S and transfected MSC_S to osteoblast and progress qualitation defection by ALP and OC then to student and reseach bone defect recovery in vivo.RESULT:1. Morphology and ultrastruture of MSCs: A samall percentage of isolated cells were adherent to the flasks and grow as typically fibroblastic or sprindle shape, which proliferated rapidly. Primary cultures reached 80-90% of confluence in about 7 to 10 days.Nonadherent or loosely adherent small round cells were present ound cells in primary cultures.2. Osteogensis Differentiation: When MSCs were grown in osteogensis inducing medium,the cells showed typical cuboidal shape. The trial groups stably expressed Alkaline Phosphatase while the controlled group remained at low level.