CAJ | 학술논문

目的应用改良植块法体外培养大鼠阴茎海绵体平滑肌细胞.方法 15只SD雄性大鼠,随机分成3组,每组5只,分别采用组织块法、酶消化法及改良组织块法分离大鼠阴茎海绵体平滑肌细胞,在含双抗及20%胎牛血清DMEM,37℃、5%CO2.95%空气的细胞培养箱静置中培养,相差显微镜观察细胞形态及扩增情况,用α-平滑肌肌动蛋白(a-SM-Actin)及结蛋白(desnun)免疫组织化学染色,鉴定细胞类型.结果 α-平滑肌肌动蛋白在阴茎海绵体平滑肌细胞阳性率为96.3%.在成纤维细胞中阳性率为23.8%,结蛋白在阴茎海绵体平滑肌细胞阳性率为74.4%,在成纤维细胞中呈阴性.三组间结蛋白阳性细胞率具有显著差异,改良组织块法组结蛋白阳性细胞率高于组织块法和酶消化法组.结论结蛋白是鉴定海绵体平滑肌细胞的特异性指标,改良组织块法可获纯度更高、结构和功能良好的阴茎海绵体平滑肌细胞.
목적 응용개량식괴법체외배양대서음경해면체평활기세포.방법 15지SD웅성대서,수궤분성3조,매조5지,분별채용조직괴법、매소화법급개량조직괴법분리대서음경해면체평활기세포,재함쌍항급20%태우혈청DMEM,37℃、5%CO2.95%공기적세포배양상정치중배양,상차현미경관찰세포형태급확증정황,용α-평활기기동단백(a-SM-Actin)급결단백(desnun)면역조직화학염색,감정세포류형.결과 α-평활기기동단백재음경해면체평활기세포양성솔위96.3%.재성섬유세포중양성솔위23.8%,결단백재음경해면체평활기세포양성솔위74.4%,재성섬유세포중정음성.삼조간결단백양성세포솔구유현저차이,개량조직괴법조결단백양성세포솔고우조직괴법화매소화법조.결론 결단백시감정해면체평활기세포적특이성지표,개량조직괴법가획순도경고、결구화공능량호적음경해면체평활기세포.
Objective To culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method.Methods Fifteen male rats were randomized into 3 equal groups, namely enzyme digestion group, tissue culture group, and modified tissue culture group. The penis of the rats was separated carefully and cut into small pieces, and seeded onto culture flasks and cultured in complete medium consisting of DMEM containing 20% fetal calf serum at 37 ℃ in a humidified atmosphere with 5% carbon dioxide. The cells growth was observed under phase contrast microscope and the smooth muscle cell specific proteins α-SM-actin and desmin were identified immunohistochernically. Results The α-SM-actin-positive cell rate was 96.3% in rat corpus cavernosum smooth muscle and 23.8% in the fibroblasts, and the corpus cavernosum smooth muscle contained 74.4% desmin-positive cells while the fibroblasts showed no desmin positivity. Significant difference was found in the positive cell rate for desmin among the 3 groups, with the highest positive cell rate occurred in modified tissue culture group. Conclusion Desmin may serve as a marker for identifying corpus cavernosum smooth muscle cells. The modified tissue culture method can result in highly purified corpus cavernosum smooth muscle cells with intact structure and functions.