ObjectiveTo explore the effects of several immunosuppressants on the proliferation of pheochromocytoma 12 (PC12) and L929 cells. 〖WTHZ〗Methods Different concentrations of methylprednisolone(10-3,10-4, 10-6and 10-8 mol/L), cyclosporin A(CsA,10-5 ,10-6 , 10 -7and 10-8 mol/L) and FK506 (10-6 ,10-7 , 10-8and 10-9mol/L)were administrated to the PC12 and L929 cells, while control group was given no drugs. At 24, 48 and 72 hours after administration, the cell proliferationwasmeasured with MTT methods respectively. The results were compared and analyzed statistically. Results High concentration methylprednisolone (10-3 mol/L) and low concentration CsA (10-8-10-7mol/L) could promote the proliferation of PC12 cells within 48 hours after administration, after that, the proliferation effects were no longer significant. There were no promotion effects for different concentrations of FK506. Under high concentrations, both CsA (10-6 -1×10-5 mol/L) and methylprednisolone (10-3 mol/L) could significantly inhibit the proliferationof L929 cells after 24 hours of administration. And high concentration (10-6mol/L) FK506 could promote the proliferation of L929 cells transitorily (only for 48 hours after administration). Conclusion 10-3 mol/L methylprednisolone and 10-8 -10-7mol/L CsA can promote the proliferation of PC12 cells for a short period of time. Both 10-3 mol/L methylprednisolone and 10-6-10-5mol/L CsA can significantly inhibit proliferation of L929.
OBJECTIVE: To study the technique and method of urethral epithelium culture in vitro, so as to lay the groundwork for reconstructing a tissue engineering urethra and to provide an experimental model of urethral mucosa in physiological, pathological, toxicological and microbiological study. METHODS: The urethral mucosa from a young male New Zealand hare that had just been out of milk, was digested into single cell liquid with Dispase II and mixed enzyme, and the fibroblast were removed. After being seeded, the cells were cultured by using L929 cells as trophoderm. The medium was changed regularly and the cells were subcultured when they grew to mix together 80% to 90%. The cultured cells were analyzed with histochemistry, immunohistochemistry dyeing and flow cytometry examination. We observed the ultrastructure of cells with scanning electron microscope and transmission electron microscope. RESULTS: The primary cultured cells fused when they had been cultured for about ten days. They were the same in size like road rocks. The cultured cells were all epithelial cells without fibroblasts and were diploid cells. The cells could be subcultured 11-13 generations, and could survive 50-60 days. CONCLUSION: The urethral epithelium of young New Zealand hare can be cultured in vitro and maintain the ability to proliferate within a certain time. The study result not only sets a role in reconstructing a tissue engineering urethral mucosa, but also provides an experimental model for the research of urethral mucosa in vitro.