大鼠肺毛细血管周细胞的培养  

Culture of rat pulmonary capillary pericytes

作　　者：陈兵[1] 易斌[1] 杨勇[1] 王芝[1] 陈杨[1] 祖宝利[1] 鲁开智[1] 

机构地区：[1]第三军医大学西南医院麻醉科,重庆市400038

出　　处：《中华麻醉学杂志》2013年第10期1242-1244,共3页Chinese Journal of Anesthesiology

摘　　要：目的 建立可靠、简便的肺毛细血管周细胞的培养方法.方法 选择健康雄性SD大鼠6只,6～7周龄,体重200 ～ 220 g,采用Ⅰ型胶原酶消化法和微孔过滤法分离肺毛细血管片段,用含10％胎牛血清和0.5％青霉素链霉素混合液的高糖DMEM/F12 1∶1培养基选择性培养肺毛细血管周细胞.倒置相差显微镜下观察细胞形态和生长状态,采用免疫荧光法计数α-平滑肌肌动蛋白（α-SMA）、结蛋白、神经元-胶质抗原2（NG2）、分化抗原31（CD31）的阳性细胞,计算阳性细胞百分比.结果 镜下细胞呈梭型或星芒状,单核,偶见双核,核卵圆形,细胞浆丰富,漩涡或栅栏状生长,无接触性抑制.免疫荧光测定α-SMA阳性细胞百分比为（99.0±1.2）％,结蛋白阳性细胞百分比为（96.0±2.1）％,NG2阳性细胞百分比为（99.0±0.7）％,CD31阳性细胞百分比为0.结论 成功建立了一种可靠、简便的大鼠肺毛细血管周细胞的培养方法.Objective To establish a method for culture of rat pulmonary capillary pericytes.Methods Six male Sprague-Dawley rats,aged 6-7 weeks,weighing 200-220 g,were anesthetized and the chest was opened.The pulmonary capillary was isolated by type Ⅰ collagenase digestion and micropore filtration and cultured in highglucose DMEM/F12 1∶1 containing 10％ fetal bovine serum and 0.5％ mixture of penicillin and streptomycin.The morphology and growth of cells were observed with inverted phase contrast microscope.The positive cells of αsmooth muscle actin （α-SMA）,desmin,neuron-glial antigen 2 （NG2）,cluster of differentiation 31 （CD31） were counted by immunofluorescence.The percentage of positive cells was calculated.Results The microscopic examination showed cells of shuttle shape or star shape,mononuclear cells,binuclear cells occasionally,oval nucleus,rich cell plasma,growth in the shape of vortex or fence,and no contact inhibition.The percentage of positive cells ofα-SMA,desmin,NG2,and CD31 was （99.0± 1.2）％,（96.0±2.1）％,（99.0±0.7）％ and0,respectively.Conclusion The culture method for rat pulmonary capillary pericytes is successfully established.

关 键 词：周细胞 细胞培养技术 肺 微脉管 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]