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目的 观察接种密度和α 二氟甲基鸟氨酸 (DFMO)对小肠上皮细胞 (IEC 6 )体外增殖的影响。方法 IEC 6细胞以不同密度接种于 96孔培养板 ,加入或不加DFMO。细胞接种后第 1,2 ,3,4 ,5 ,6和 7天 ,分别用细胞计数板直接计数细胞数量 ,用噻唑蓝比色法 (MTT法 )测定波长 5 70nm处的光密度 (OD)值 ,确定细胞的增殖活力。结果 OD值和细胞数量具有正相关 (r=0 95 4,P <0 0 1) ;较高接种密度 (>0 5× 10 4 细胞 /孔 )组接种后第 2天 ,细胞生长抑制 ,尤其是密度增加到 4× 10 4 细胞 /孔时 ,第 2～第 3天OD值逐渐增加 ,第 4天明显增加 ,第 5天达高峰 ,此后OD值开始下降。而低密度 (0 2× 10 4 细胞 /孔 )接种后第 4天 ,细胞生长出现抑制 ,此后其生长与其它密度相似。另外 ,加入DFMO后 1～ 3天 ,完全抑制了细胞增殖 ,此后与对照组一样 ,细胞逐渐生长。结论 IEC 6细胞增殖与接种密度和培养时间有关 ;随着培养时间的延长 ,DFMO抑制细胞增殖作用具有可逆性 ,其与培养时间延长有关。
Objective To observe the effects of inoculation density and α-difluoromethylornithine (DFMO) on the proliferation of small intestinal epithelial cells (IEC 6) in vitro. Methods IEC 6 cells were seeded on 96-well plates at various densities with or without DFMO. The number of cells was counted directly on the 1st, 2nd, 3rd, 4th, 5th, 6th and 7th day after inoculation. The optical density (OD) at 570nm was measured by MTT method. Value, determine the cell proliferation activity. Results There was a positive correlation between OD value and cell number (r = 0 954, P <0.01). On the 2nd day after inoculation with higher seeding density (> 0 5 × 10 4 cells / well), cell growth was inhibited When the density increased to 4 × 10 4 cells / well, the OD values gradually increased from the second day to the third day, increased significantly on the fourth day and reached the peak on the fifth day, and the OD value began to decline thereafter. On the 4th day after inoculation at a low density (0 2 × 10 4 cells / well), cell growth was inhibited and its growth thereafter was similar to other densities. In addition, after adding DFMO 1 to 3 days, completely inhibited cell proliferation, then as the control group, the cells gradually grow. Conclusion The proliferation of IEC 6 cells is related to the inoculation density and culture time. With the prolongation of culture time, the effect of DFMO on cell proliferation is reversible, which is related to the prolongation of culture time.