CAJ | 학술논문

背景:肾小球系膜细胞原代培养成功率低,生存时间短,传代次数少.其中分离提取肾小球一直是培养高纯肾小球系膜细胞的最大瓶颈.目的:建立一种操作简单、成功率高、重复性好的人肾小球系膜细胞原代培养方法.方法:取自愿水囊引产的胎儿肾,采用肾皮质组织块法合优生选择法培养人肾小球系膜细胞.相差显微镜、透射电镜观察系膜细胞形态,免疫荧光技术鉴定细胞表型,激光共聚焦显微镜观察波形蛋白的表达.结果与结论:培养的肾小球系膜细胞呈梭形/不规则星形和细长形,胞质中各种细胞器丰富,细胞突起明显,胞膜上有很多微绒毛,细胞表达α-肌动蛋白、肌球蛋白、波形蛋白、结蛋白,而不表达细胞角蛋白、Ⅷ因子,激光共聚焦显微镜下发现系膜细胞波形蛋白表达阳性并具有特征性纤维束.说明培养的系膜细胞符合肾小球系膜细胞的生物学特征.肾皮质组织块法合优生选择法是一种简单、高效的培养人肾小球系膜细胞的方法.
배경:신소구계막세포원대배양성공솔저,생존시간단,전대차수소.기중분리제취신소구일직시배양고순신소구계막세포적최대병경.목적:건립일충조작간단、성공솔고、중복성호적인신소구계막세포원대배양방법.방법:취자원수낭인산적태인신,채용신피질조직괴법합우생선택법배양인신소구계막세포.상차현미경、투사전경관찰계막세포형태,면역형광기술감정세포표형,격광공취초현미경관찰파형단백적표체.결과여결론:배양적신소구계막세포정사형/불규칙성형화세장형,포질중각충세포기봉부,세포돌기명현,포막상유흔다미융모,세포표체α-기동단백、기구단백、파형단백、결단백,이불표체세포각단백、Ⅷ인자,격광공취초현미경하발현계막세포파형단백표체양성병구유특정성섬유속.설명배양적계막세포부합신소구계막세포적생물학특정.신피질조직괴법합우생선택법시일충간단、고효적배양인신소구계막세포적방법.
BACKGROUND:Primary culture of glomerular mesangial cells was less achievement ratio,short survival time,and less passage times.In particular,extraction of renal glomerulus remains difficult for culturing highly pure mesangial cell OBJECTIVE:To establish a more simple.high successful rate and good reproducibility method of human mesangial cells in primary cultureMETHODS:Kidneys jsolated from induction of labor with water bag voluntary were cut into pieces.and human mesangial cells were cultured with eugenic selection methods.Morphology was observed using inverted phase contrast microscope and transmission electron microscope,cell phenotype was detected using immunohistochemical method,and vimentin expression was observed using laser scanning confocal microscope.RESULTS AND CONCLUSION:The cultured mesangial cells were fusiform-shaped,irregular star-shaped,and slender.Organelle was rich in cytoplasm,cell process was clear,and microvillus was observed on the cell membrane.The cells expressed a-actin,myosin,vimentin,desmin but not expressed cytokeratin and Ⅷ factor.Laser scanning confocal microscope demonstrated that vimentin expression was positive and had the characteristics of fiber bundles.This suggested that the cultured intercapillary cells were coincidence with the characteristics of mesangial cell The renal corticaI tissue combined eugenic selection method was a simple and efficient method to culture human mesangial cells.