Study of newly autologous chondrocyte implantation using by gene transfer of hyaluronan synthase

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In the present study, plasmids containg HAS2 under the PMX-IRES-GFP retrovirus vector were transfected into undifferentiated ATDC5. As a control, cells transfected with the empty vector were used. ATDC5 cells differentiated in the presence of insulin and RT-PCR was performed to detect HAS2 m RNA expression and we evaluated visualization of pericellular coats which were composed of newly synthesized HA using by immunofluorescent staining and analyzed HA concentration in culture medium by high performance liquid chromatography. The signal of the cells transfected with HAS2 was visually much stronger than that of control cells. The HA concentration of the cells transfected with HAS2 in culture medium was much higher than that of control throughout 21 days. By transfection of HAS2 with retrovirus, we are able to expect the continuous synthesis of HA. Therefore this technique may be a great significance in autologous chondrocyte implantation.