CAJ | 학술논문

目的建立成人主动脉血管平滑肌细胞体外培养的有效方法.方法无菌条件下分离成人主动脉的平滑肌层,剪成1 mm3的碎片,0.1％ Ⅰ型胶原酶预处理1h,组织块贴壁法,以含胎牛血清20％的DMEM低糖培养基培养.倒置显微镜观察培养细胞的形态学特点,免疫荧光法检测平滑肌细胞肌动蛋白(α-SMA)的表达.结果培养至4～5d组织块边缘即有少量细胞爬出,呈长梭形,胞质丰富；培养至1～2周组织块边缘细胞融合,呈典型的“峰谷”样表现.免疫荧光染色结果显示胞质内α-SMA的表达丰富,荧光强度在(++)～ (+++).结论胶原酶预处理组织的改良组织块贴壁法培养人主动脉血管平滑肌细胞是一种可行有效的实验方法.
목적 건립성인주동맥혈관평활기세포체외배양적유효방법.방법 무균조건하분리성인주동맥적평활기층,전성1 mm3적쇄편,0.1％ Ⅰ형효원매예처리1h,조직괴첩벽법,이함태우혈청20％적DMEM저당배양기배양.도치현미경관찰배양세포적형태학특점,면역형광법검측평활기세포기동단백(α-SMA)적표체.결과 배양지4～5d조직괴변연즉유소량세포파출,정장사형,포질봉부；배양지1～2주조직괴변연세포융합,정전형적“봉곡”양표현.면역형광염색결과현시포질내α-SMA적표체봉부,형광강도재(++)～ (+++).결론 효원매예처리조직적개량조직괴첩벽법배양인주동맥혈관평활기세포시일충가행유효적실험방법.
Objective To establish an effective method of human vascular smooth muscle cells in vitro culture.Methods Medial layer of human aorta was separated under sterile conditions,then cut into small pieces (about 1 mm3).After pretreatment with 0.1％ type Ⅰ collagenase,the tissues were transferred into flask and cultured in DMEM containing 20％ fetal bovine serum,penicillin (100 U/ml) and streptomycin (100 g/L).The cell morphology was observed under the inverted microscopy.Smooth muscle cell specific protein (α-SMA) was identified by using immumofluorescence methods.Results Some smooth muscle cells started growing from explants in 4 to 5 days.One to 2 weeks later,the cells grew to confluence by migrated cells.Immumofluorescence staining showed positive expression of α-SMA (fluorescence intensity (++)-(+++)).Conclusion Modified explant technique by pretreating tissues with collagenase was an effective method to get vascular smooth muscle cells in vitro.