与人脐动脉平滑肌细胞共培养的人脐静脉内皮细胞的生物学特性

Biological characteristics of human umbilical venous endothelial cells co-cultured with human umbilical arterial smooth muscle cells

目的:通过共培养人脐静脉内皮细胞(human umbilical venous endothelial ceils,HUVEC)与人脐动脉平滑肌细胞(hunlan umbilical arterial smooth muscle cells,HUASMC),初步探讨观察共培养的HUVEC的形态和增殖特性及经皮冠状动脉成形术(percutaneous transluminal coronary angioplasty,PTCA)后再狭窄的机制。 方法:用共培养装置将体外同时培养在一个培养孔中的HUVEC和HUASMC分离。在倒置显微镜下观察单独培养和共培养HUVEC的生长状态和形态;采用免疫荧光法标记细胞,流式细胞仪测定荧光强度和细胞周期。 结果:共培养的HUVEC逐渐转变为长梭形;单独培养的HUVEC仍保持鹅卵石样。单独培养的和共培养的HUVEC处于细胞周期GO/G1期和G_2+s期的百分率为:(70±3)％和(81±5)％:(22±4)％和(14± 4)％。二者均有Cyclin D的表达,但后者的表达显著低于前者。结论:共培养的HUVEC低血清干预48h形态逐渐变为长梭形,更接近于在体形状;而单独培养的HUVEC仍保持原鹅卵石状。此外,前者的增殖活性也显著低于后者。

AIM: By co-culturing human umbilical arterial smooth muscle cells (HUASMC) with human umbilical venous endothelial cells (HUVEC), to explore the morphological and proliferation characteristics of HUVEC, and the mechanism of restenosis after percutaneous transluminal coronary angio-plasty(PTCA). METHODS: Using the co-culture dish, HUVEC were separated with HUASMC, which were cultured in the same porus in vitro. The growth behavior and morphologic changes in both single cultured and co-cultured HUVEC were observed by invert microscope, cells were labeled with im-munofluorescence, and fluorescent intensity and cell cycle were inspected by flow cytometer. RESULTS: The co-cultured HUVEC went longer, like fusiform, but the single cultured remained cobble . The percent of the single cultured and co-cultured HUVEC in G0/G1 or G2 + S period were (70 ±3)% and(81 ± 5)%; (22 ±4)% and(14±4)%. The expression of cyclin D was significantly lower in co-cultured HUVEC than in single cultured HUVEC. CONCLUSION: The single cultured HUVEC remains 'cobble', but the co-cultured goes fusiform after 48 h lower serum intervention and are similar to those in vivo. In addition, proliferation of the former is significantly lower than that of the later.