创面收集液对表皮干细胞表型的影响  

Effects of wound harvested effusion on phenotype of epidermal stem cells in vitro

作　　者：向启利[1] 李建福[1] 付小兵[2] 

机构地区：[1]第三军医大学附属西南医院整形科,重庆400038 [2]解放军总医院基础部

出　　处：《中华创伤杂志》2007年第6期452-455,共4页Chinese Journal of Trauma

基　　金：国家自然科学基金面上资助项目（30570708）;国家重点基础研究发展规划资助项目（2005CB522603）

摘　　要：目的 探讨全层创面收集液对表皮干细胞类型的影响。方法 以聚氨酯海绵收集成年Wistar大鼠背部全层创面渗出液；以快速黏附分离法体外分离、培养新生Wistar大鼠的表皮干细胞，待表皮干细胞呈克隆生长时加入创面收集液干预，分别于干预后的0，6，12，18，24，30，36，42，48，54，60，72h采用流式细胞仪及β1整合素、角蛋白19，14，10免疫组化染色进行细胞增殖分化的鉴定。结果表皮干细胞在创面收集液干预后仍可持续保持片状聚集生长的态势，但出现数量较多的角蛋白14阳性染色细胞群落，且随时间延长该阳性细胞群落呈上升趋势；另出现了散在的角蛋白10染色阳性细胞。结论创面收集液可以快速诱导体外培养表皮干细胞进入分化状态，且多表现为短暂扩充细胞表型，在该过程中表皮干细胞数量仍可维持在一定的水平。Objective To investigate the effects of wound harvested effusion （WHE） on phenotype of epidermal stem cells （ESCss）. Methods Wound effusion was harvested with polyurethane sponge from 80 full-thickness wounds on the backs of 40 adult Wistar rats. ESCss were isolated, purified fi＇om neonatal Wistar rats and then treated with the wound effusion when they showed clone growing. 131 integrin, keratin 19 （K19） , Keratin 14 （K14） and Keratin 10 （K10） were employed to determine differentiation status of ESCss with immunohistochemical staining methods and flow cytometer technology at 0, 6^th, 12^th, 18^th, 24^th, 30^th, 36^th, 42^nd, 54^th, 60^th, 72^nd hours after treatment. Results ESCss could still keep growth in colony after treatment with WHE. There were a number of K14 positive cell populations that gradually increased in number with treating time. However, a few positive of K10 stained cells were found scattering within the K14 positive cell populations. Conclusion WHE can directly induce ESCss to be under differentiation, when the differentiated cells phenotype displays the feature of transient amplifying cells and keeps certain number of ESCss.

关 键 词：干细胞 表皮 细胞分化 表型 渗出液和漏出液 创面 

分 类 号：R686[医药卫生—骨科学]