大鼠肾小球系膜细胞体外培养及鉴定的实验研究  

Experimental Study of Glomerular Mesangial Cell from Rats in vitro and Its Identification

作　　者：翟文生[1] 白洁[1] 王文英[1] 郭会敏[1] 刘华[2] 平东沛[1] 赵小静[1] 高旭光[1] 

机构地区：[1]河南中医学院第一附属医院,河南郑州450008 [2]河南中医学院,河南郑州450008

出　　处：《中医学报》2011年第3期319-321,共3页Acta Chinese Medicine

基　　金：河南省杰出青年计划(编号:084100510018)

摘　　要：目的:探索一种重复性好、传代次数多的大鼠肾小球系膜细胞(glomerular mesangial cell,GMC)体外培养和鉴定的方法。方法:麻醉状态、无菌条件下取得大鼠的肾脏,用3种不同目数的不锈钢筛网提取肾小球,搜集到的肾小球分别加入2.5 g/L、0.5 g/LⅣ型胶原酶及2 g/LⅡ型胶原酶各5 mL,分别在37℃水浴箱中振荡消化20 min、30 min、25min,然后接种肾小球,比较不同培养条件下肾小球系膜细胞的生长状况。结果:Ⅱ型、Ⅳ型胶原酶3种不同浓度作用不同的消化时间,细胞的生长速度及生长状况不同,Ⅳ型胶原酶(2.5 g/L)消化20 min的为最佳。结论:培养的细胞为肾小球系膜细胞,用Ⅳ型胶原酶(2.5 g/L)消化20 min的肾小球系膜细胞生长状态最佳。Objective：Find a good method high specifity and subculture ability for Glomerular mesangial cells（GMC） of rat cultivated in vitro and cells characterized,the optimum conditions for culturing of GMC were summaried.Methods：Under anaesthetic and sterile condition out the kidney of rats,With three different accounts of the stainless steel micropore filter extraction glomerular,The 5ml reagents of 2.5 g/L,0.5 g/L type IV collagenase and 2 g/L type Ⅱ collagenase were respectively droped in the three tubes Collected glomerular cells.The three centrifuge tubes GMC oscillating in 37 ℃ water tank 20,30 and 25 min,Then inoculate glomerular,To compare the different conditions of the GMC grow conditions.Results：The cell growth rate and growth condition was different,when the three different concentrations of Ⅱtype collagenase and Ⅳtype collagenase digest at different times.The best condition was Ⅳ type collagenase（2.5 g/L）digested 20min.GMC growth feature were dynamicly observed under invert microscope and photographs.The cells were GMC which we cultured was identificated by morphology and immunohistochemical methods.Conclusions：The results suggest that this cell culture method was simple and efficient,the glomerular mesangial cells（GMC） of rat cultivated in vitro was successful.

关 键 词：肾小球系膜细胞 细胞培养 细胞鉴定 Ⅱ型胶原酶 Ⅳ型胶原酶 

分 类 号：R587.24[医药卫生—内分泌]