军事医学科学院院刊
軍事醫學科學院院刊
군사의학과학원원간
BULLETIN OF THE ACADEMY OF MILITARY MEDICAL SCIENCES
2010年
1期
37-39
,共3页
程孝中%宋婷%黄蓓%钟辉
程孝中%宋婷%黃蓓%鐘輝
정효중%송정%황배%종휘
人胚胎血管平滑肌细胞%原代培养%钙化模型
人胚胎血管平滑肌細胞%原代培養%鈣化模型
인배태혈관평활기세포%원대배양%개화모형
human embryo vascular smooth muscle cells%primary culture%calcification mode
目的 利用β-甘油磷酸盐处理人血管平滑肌细胞(human vascular smooth muscle cells, HVSMCs )制备体外血管钙化模型.方法 用组织贴壁法从人胚胎脐带动脉中分离原代人主动脉平滑肌细胞,原代细胞通过抗体α-sm-actin 染色鉴定,将传4~6代的细胞分为钙化组和对照组,对照组用正常DMEM培养基培养,钙化组加入10 mmol/L β-甘油磷酸盐诱导细胞钙化,连续培养10 d.茜素红S染色及碱性磷酸酶法鉴定.结果 :分离的原代细胞经S-P染色鉴定为阳性,呈淡黄色.钙化组细胞诱导钙化后,细胞增殖缓慢,并形成囊泡结构,茜素红S染色形成红色的钙化结节,钙化组碱性磷酸酶活性较对照组在不同时间点(4,6,8,10 d)都有所增加(P＜0.01).结论 :β-甘油磷酸盐体外能够诱导HVSMC钙化,此方法诱导的钙化模型是一种良好的研究血管疾病方面的体外模型.
目的 利用β-甘油燐痠鹽處理人血管平滑肌細胞(human vascular smooth muscle cells, HVSMCs )製備體外血管鈣化模型.方法 用組織貼壁法從人胚胎臍帶動脈中分離原代人主動脈平滑肌細胞,原代細胞通過抗體α-sm-actin 染色鑒定,將傳4~6代的細胞分為鈣化組和對照組,對照組用正常DMEM培養基培養,鈣化組加入10 mmol/L β-甘油燐痠鹽誘導細胞鈣化,連續培養10 d.茜素紅S染色及堿性燐痠酶法鑒定.結果 :分離的原代細胞經S-P染色鑒定為暘性,呈淡黃色.鈣化組細胞誘導鈣化後,細胞增殖緩慢,併形成囊泡結構,茜素紅S染色形成紅色的鈣化結節,鈣化組堿性燐痠酶活性較對照組在不同時間點(4,6,8,10 d)都有所增加(P＜0.01).結論 :β-甘油燐痠鹽體外能夠誘導HVSMC鈣化,此方法誘導的鈣化模型是一種良好的研究血管疾病方麵的體外模型.
목적 이용β-감유린산염처리인혈관평활기세포(human vascular smooth muscle cells, HVSMCs )제비체외혈관개화모형.방법 용조직첩벽법종인배태제대동맥중분리원대인주동맥평활기세포,원대세포통과항체α-sm-actin 염색감정,장전4~6대적세포분위개화조화대조조,대조조용정상DMEM배양기배양,개화조가입10 mmol/L β-감유린산염유도세포개화,련속배양10 d.천소홍S염색급감성린산매법감정.결과 :분리적원대세포경S-P염색감정위양성,정담황색.개화조세포유도개화후,세포증식완만,병형성낭포결구,천소홍S염색형성홍색적개화결절,개화조감성린산매활성교대조조재불동시간점(4,6,8,10 d)도유소증가(P＜0.01).결론 :β-감유린산염체외능구유도HVSMC개화,차방법유도적개화모형시일충량호적연구혈관질병방면적체외모형.
Objective To establish a calcification mode in vitro of human vascular smooth muscle cells (HVSMCs) induced by β-GP. Methods Primary HVSMCs were obtained from human embryo by plant method and confirmed by stain with α-sm-actin antibody. The cells after 4-6 passages were divided into two groups.The control group was incubated with normal DMEM medium while the calcification group was incubated with the medium containing 10 mmol/L β-glycerophosphate for 10 days.Calcification was confirmed by alizarin red S stain and alkaline phosphatase(ALP) assays. Results The primary cells observed by S-P stain were positive and the cells after being stained were pale yellow. After being induced with β-GP, the cells of calcification group began concentric growth and formed vesicles. Alizarin red S staining showed that the reaction of calcifying nodules was red,ALP activity was higher than that of controls at various time points(4 d,6 d,8 d and 10 d,P＜0.01 ).Conclusion The HVSMCs could be induced into calcification in vitro by β-GP, and this model contribates to further studies of vascular diseases.
