无血清培养基培养人毛乳头细胞的研究  

Study on serum-free culture of dermal papillae cells of human hair

作　　者：夏汝山[1] 郝飞[1] 杨希川[1] 宋志强[1] 钟白玉[1] 尹锐[1] 

机构地区：[1]第三军医大学西南医院皮肤科,重庆400038

出　　处：《中华皮肤科杂志》2006年第6期335-337,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金(30300180)

摘　　要：目的探讨无血清培养液培养人头皮毛乳头细胞(dermal papillae cells,DPC)的可行性,并研究其生长特性。方法预先应用纤维连接蛋白处理细胞培养瓶(板)；采用Williams E无血清培养液并补充ITS(牛胰岛素、转铁蛋白和亚硒酸盐)培养第2代DPC,倒置相差显微镜下观察细胞生长状态,采用MTT评价其增殖状态并绘制其生长曲线。结果在无血清培养条件下,DPC在2～4h内贴壁。2～3d后细胞可相互融合．未见细胞明显增殖；培养4d后细胞连接开始中断。形成孤立的细胞或细胞团,并可见细胞的脱落；培养1～2周后,细胞大多脱落。给予含4%小牛血清的DMEM培养10h后,在残余的细胞团周围长出放射状排列的细胞；生长曲线显示DPC的生长呈现停滞期和缓慢生长期,未见指数生长期。结论采用无血清培养基可以培养DPC,但其培养条件有待优化。Objective To explore the feasibility of culturing dermal papillae cells （ DPC ） of human hair in a serum-free medium, and to observe the growth characteristics of these cells. Methods Cell culture flasks （plates） were pretreated with fibronectin, and DPC （2nd passage ） were incubated with Williams E serum-free medium supplemented with insulin-transferrin-selenite （ ITS ）. Cells were observed by an inverted phase-contrast microscope. Proliferation of DPC was evaluated with 3- （ 4,5-dimethylthia- zol-2-yl ）-2,5-diphenyl tetrazolium bromide （MTT） assay and by their growth curve. Results In a serum-free medium, 2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later, confluence, of the cells was observed, without noticeable proliferation. Four days later, cell connection was interrupted, isolated cells or cell clusters were seen, and detachment of some cells from the flask surface was observed. One to two weeks later, most cells had died. After incubation with 4% bovine serum for ten hours, cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase; however, log growth phase was not observed. Conclusion DPC could be successfully cultured in serum-free medium. However, the culture condition needs to be further optimized.

关 键 词：培养基 无血清 毛乳头 细胞培养 

分 类 号：R751[医药卫生—皮肤病学与性病学]