大鼠肺微血管内皮细胞培养及其粘弹性研究  

Investigation on Culture of Rat Pulmonary Microvascular Endothelial Cells and Their Viscoelasticity

作　　者：李发琪[1] 杨瑞芳[2] 黄岂平[2] 秦健[2] 蔡绍皙[2] 吴云鹏[2] 

机构地区：[1]重庆医科大学医学超声工程研究所,重庆400016 [2]重庆大学生物工程学院生物力学&组织工程教育部重点实验室,重庆400044

出　　处：《生物医学工程学杂志》2002年第1期36-39,共4页Journal of Biomedical Engineering

摘　　要：为了建立肺微血管内皮细胞培养方法 ,研究肺微血管内皮细胞粘弹性。我们取大鼠肺周边组织 (宽度不应大于 1.5 mm) ,将组织剪成 1.5 mm× 1mm× 1mm的组织块 ,贴入无菌的 2 5 cm3培养瓶 ,每瓶 10～ 15块 ,同时加入含 2 0胎牛血清、肝素 90 U/ml、L-谷氨酰胺 4mmol、青霉素 10 0 U/ml和链霉素 10 0 μg/ml的 DMEM培养基 3ml,放入 37℃二氧化碳培养箱中静置培养 ;8h后翻转培养瓶 ,6 0 h后取出肺组织块 ,接着继续培养 2～ 4d后进行传代。最后消化分离肺微血管内皮细胞 ,用微管吸吮系统研究肺微血管内皮细胞粘弹性。结果显示 :肺微血管内皮细胞通过倒置相差显微镜观察 ,细胞呈鹅卵石镶嵌状排列 ,状如梭形或多角形 ,大小均匀 ,胞核清晰 ,呈卵圆形 ,胞浆丰富 ; 因子相关抗原免疫荧光染色呈阳性 ;肺微血管内皮细胞弹性模量 K1 =49.3± 9.2 Pa、K2 =73.2±2 4.8Pa、粘性系数 μ=19.2± 7.2 Pa.s。这些结果表明用组织块法培养肺微血管内皮细胞是可行的 。It is the infent of this study to establish a simple method for cultivation of rat pulmonary microvascular endothelial cells(PMVECs) and investigate the viscoelasticity of PMVECs. First, we obtained rat's peripheral pulmonary tissue,which then was cut into small pieces and cultured with 3ml DMEM containing 20% bovine calf serum ,90U/ml heparin, 4mmol L glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin. Next, moved away the pulmonary tissue pieces 60h later,and started passage 2 4 days after continued culture. Last, digested and separated PMVECs and studied viscoelastic coefficients of PMVECs by using micropipette aspiration technique. The results revealed that the cultured PMVECs showed regular cobblestone morphology and conformed with endothelial cells morphological characterization by phase contrast microscopy. PMVECs elastic modulus K 1 was 49.3±9.2 Pa,K 2 was 73.2±24.8 Pa, and it's viscosity factor μ was 19.2±7.2 Pa.s. These data demonstrate that it is feasible to cultivate PMVECs with tissue pieces method,and PMVECs is of greater rigidity.

关 键 词：肺微血管内皮细胞 细胞培养 微管吸吮技术 细胞粘弹性 

分 类 号：R318.01[医药卫生—生物医学工程]