乳大鼠表皮细胞体外培养及环孢素A对其增殖抑制作用

Culture of epidermal cells and inhibition effect of cyclosporin A on cell proliferation in infant mice

目的观察无表皮生长因子(EGF)条件下体外培养乳大鼠表皮细胞生长特点,及不同浓度的环孢表A(CSA)对细胞增殖的影响。方法用单细胞悬液法体外培养出生24h内的乳大鼠的表皮细胞 ,观察其生长曲线和CSA对细胞增殖的抑制作用。结果在无EGF条件下 ,细胞培养的最长时间为15天。接种24h开始贴壁 ,第2天开始生长 ,第5～7天达高峰 ,第10天开始在胞浆中出现空泡 ,以后逐渐死亡。1μg/mlCSA在24h、48h对细胞增殖无明显抑制作用 ,72h时的抑制率为15 %;3μg/mlCSA在24h也无抑制作用 ,48h和72h的抑制率分别达18 %和30 %;6μg/mlCSA培养24h、48h、72h后抑制率分别为16 %、40 %和65 %。结论无EGF时培养的乳大鼠表皮细胞在5～7天生长达高峰 ,最长培养时间为15天。CSA对细胞增殖有抑制作用 ,且有明显的浓度和时间依赖性。

Aim The growth characteristic of cultured epidermal cell and the inhibition effect of cyclosporin A (CSA) on cell proliferation at the doses of l μg/ml,3μg/ml and 6 μg/ml were observed. Methods The single epidermal cells of infant mice were cultured without EGF (eidermal growth factor). Growth curve was descriped by numbering the alive cells in different times. The inhibition rate of CSA on cell proliferation was calculated. Results Epidermal cells might be survived for about 15 days without EGF. Cells would attach to the wall of bottle in 24 h. The growth top time was in 5～7 days. There were some hollows in cytoplasm after 10 days. There was no inhibition effects of CSA on cell proliferation when cells were cultured for 24 h and 48 h at the dose of 1 μg/ml, but a weak inhibition effect was seen in 72 h, with the inhibition rate being 15% when cells were cultured with 3 μg/ml CSA for 48 h and 72 h, the inhibition rates were 18% and 30%. There was no inhibition effect in 24 h still. When cells were cultured with 6μg/ml for 24 h, 48 h and 72 h the inhibition rates were 16% ,40% and 65% respectively. Conclusion CSA inhibit cell proliferation in a dose and time dependent manner.