改良混合酶消化法培养大鼠主动脉平滑肌细胞  

Improvement of Cultured Rat Vascular Smooth Muscle Cells by Enzymatic Dispersion Method

作　　者：李成武[1,2] 李应续[2] 朱从丽[1,3] 宋健[1,3] 

机构地区：[1]武汉大学基础医学院人体解剖学与组织胚胎学系,湖北武汉430071 [2]咸宁学院基础医学院人体解剖学教研室 [3]武汉大学基础医学院医学研究中心

出　　处：《咸宁学院学报（医学版）》2008年第5期369-372,共4页Journal of Xianning Univarsity(medical Sciences)

基　　金：国家自然科学基金资助项目(30470906)

摘　　要：目的探讨改良酶消化法培养大鼠主动脉平滑肌细胞(VSMC)的方法,为动脉粥样硬化等血管硬化性疾病的发病机制和防治研究提供帮助。方法提取大鼠的胸主动脉,采用混合酶消化后,采用含10%FCS RP-M I1640培养基培养。通过细胞计数观察细胞增殖,倒置相差显微镜观察血管平滑肌细胞的生物学性状,免疫组织化学进行细胞鉴定。结果消化肌平滑肌细胞24h后贴壁,平均贴壁率为76%,第5 d后细胞数迅速增加,至第11 d形成单层时,光镜下出现"峰-谷"样生长;传代后,细胞的生长特性无改变,α-SMA免疫组化染色检测,证实细胞是VSMC。结论改良后酶消化法能短时、高效地培养生物学特征稳定的VSMC。Objective To explore the cultivation of rat vascular smooth muscle cells （VSMC） by erizymatic dispersion which helps to provide ideals for the prevention and treatment of angiosclerotic diseases. Methods The thoracic artery was immediately removed and cut into fme tissue pieces, then was incubated in a mixed medium. After digestion,the cells were cultured in RPMI1640 with 20% FCS and then plated onto plastic tissue culture dishes. The proliferation of cell was assayed by cell counting, the feature of vascular smooth muscle cells was observed with inverted phase contrast microscope, and immunohistochemical method was used to indentify cells. Results After 24h of incubation,VSMCs began to stick to the wall of the incubation dishes,and the adherent rate was 76% . After 5th day, the cell number was increased rapidly, and they form＇monolayer on 11th day. A ＂ the peak-valley＂ type of growth could be observed by light microscope. After passages, the growth characteristics of cells did not changed, and the confirmation cell was VSMC by α-SMA immunohistochemical testing. Conclusion The improved enzymatic dispersion method for VSMC can increase the cell stability of biological characteristics with short-time.

关 键 词：酶消化法 平滑肌细胞 胸主动脉 SD大鼠 血管硬化性疾病 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]