免疫磁珠分选人角朊干细胞的实验研究  

Experimental Study on Magnetic Activated Cell Sorting of Human Keratinocyte Stem Cells

作　　者：罗海水[1] 彭代智[1] 周新[1] 刘敬[1] 朱崇涛[1] 严泉[1] 李星[1] 王勇[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆400038

出　　处：《现代生物医学进展》2007年第6期812-814,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(30471679);创伤烧伤复合伤国家重点实验室开放基金课题(200306)

摘　　要：目的:探索人角朊干细胞(keratinocyte stem cells,KSCs)的免疫磁珠分选(Magnetic Activated Cell Sorting,MACS)的方法。方法:从人包皮组织获得角朊细胞悬液,然后利用人角朊干细胞表面CD49f(整合素α6)和CD71的表达情况(CD49fbriCD71dim)通过MACS法将KSCs分选出来,在荧光显微镜下观察其荧光标记情况,同时将分选所得的CD49fbriCD71dim细胞进行体外无血清培养,观察其形态及克隆形成。结果:人角朊细胞经MACS法分选后所得的CD49fbriCD71dim细胞比率可达12．02(±6．92)%。CD49fbriCD71dim细胞经培养可见细胞为典型的上皮样特征,呈铺路石样形态,高核浆比例,细胞紧密排列,轮廓清楚,折光性好,并可在5-7天左右形成全克隆,符合KSCs的特点。结论:研究表明MACS分选法可以得到较高比例的人角朊干细胞,并能进行后续培养研究,其不失为一种较理想的人角朊干细胞的分选方法。Objective： To investigate the Magnetic Activated Cell Sorting （MACS） of human keratinocyte stem cells （KSCs）. Methods： Kerafinocytes were isolated from human foreskins and then KSCs were separated by MACS according to expression of surface CD49f （integrina6） and CD71 （CD49fbriCD71dim）. The fluorescene labeling of the CD49fbriCD71dim cells was observed under fluorescene microscope. Furthermore, the morphological change and colony-forming were observed after CD49fbriCD71dim cells were cultured in serum-free medium. Results： After MACS, the ratio of CD49fbriCD71dim from kerafinocyte cells reached 12.02 （±6.92）%. Typical epithelium cells can be observed just like pebbles. These cells were dose-up and had high karyoplasmic ratio and clear circumscription. And then their colony-forming could be observed after 5-7 days. The above-mentioned was characteristic of KSCs. Conclusion： This research indicates that through MACS,a large proportion of KSCs can be obtained, and following culture researches would be making,which is indeed an ideal sorting method for KSCs.

关 键 词：角朊干细胞 免疫磁珠分选 克隆形成 

分 类 号：Q813[生物学—生物工程]