不同时间收集的KCCM对HSC-T_6细胞的增殖和胶原合成的影响

Effects of Kupffer cell-conditioned medium harvested at different time on proliferation and collagen production of HSC-T_6 cells

目的 研究不同时间收集的枯否细胞条件培养基(KCCM)对体外HSC—T6细胞的增殖和胶原合成的影响。方法 用CCl4急性损伤的大鼠肝制备KCCM,采用大鼠肝星状细胞——HSC—T6细胞株以及[3H]-TdR和[3H]-Proline掺入法分别检测其增殖和胶原合成的活性。结果 系列倍比稀释的KCCM与HSC-T6细胞共同孵育48 h后,KCCM稀释比为1:4时促增殖和胶原合成的能力最强。每隔48 h收集1次KCCM,共7次,前6次收集的KCCM能明显促进HSC-T6细胞合成胶原(P<0.01);前5次收集的KCCM能明显促进HSC-T6细胞增殖。结论 制备KCCM时,最多可收集6次。

OBJECTIVE: To study the effects of Kupffer cell-conditioned medium (KCCM) harvested at different time on proliferation and collagen production of HSC-T6 cells. METHODS: KCCM was obtained from the liver of adult rat injured by CCl4. Proliferation and collagen stimulating activity of KCCM were measured by [3H]-TdR and [3H]-Proline incorporation assay respectively using rat hepatic stellate HSC-T6 cell. RESULTS: After HSC-T6 cells were incubated with a series of KCCM diluted by double ratio for 48 h, the proliferation and collagen synthesis in HSC-T6 cells exposed to KCCM (1:4) were remarkably enhanced. KCCM harvested at first six times remarkably stimulated collagen synthesis of HSC-T6 cells (P 6 cells. CONCLUSION: KCCM may be harvested for six times at most during preparation.