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目的研究不同培养基对人脐静脉内皮细胞体外培养生长的影响,探讨内皮细胞的最佳体外扩增培养条件。方法取健康产妇分娩后脐带,用胶原酶Ⅰ消化后得脐静脉内皮细胞,进行原代培养,并用免疫荧光染色的方法对细胞进行鉴定,传代培养时则分别采用RPMI-1640培养基和EGM-2培养基,倒置显微镜观察两种培养条件下细胞的生长状态,同时利用流式细胞仪检测其生长周期,对比培养效果。结果 EGM-2组内皮细胞生长良好,2d后贴壁生长的细胞可达90%。EGM-2组S期细胞比例为(29.07±1.48)%,RPMI-1640培养基组S期细胞为(17.58±3.49)%,二者比较差异有统计学意义(P<0.01)。结论 EGM-2培养基更适合人脐静脉内皮细胞的体外传代扩增培养。
Objective To study the effects of different culture medium on the growth of human umbilical vein endothelial cells in vitro and to explore the best culture conditions of endothelial cells in vitro. Methods The umbilical cord of healthy pregnant women after delivery was harvested and the umbilical vein endothelial cells were digested with Collagenase I. The cells were primary cultured and identified by immunofluorescence staining. When subcultured, RPMI-1640 medium and EGM- 2 medium, the growth of cells under two culture conditions was observed under an inverted microscope, and the growth cycle was detected by flow cytometry to compare the culture effects. Results The endothelial cells in EGM-2 group grew well, and the adherent cells grew up to 90% after 2 days. The proportion of S phase cells in EGM-2 group was (29.07 ± 1.48)%, while that in RPMI-1640 medium group was (17.58 ± 3.49)%. The difference was statistically significant (P <0.01). Conclusion EGM-2 medium is more suitable for in vitro proliferation of human umbilical vein endothelial cells.