中文摘要：

目的：观察表皮干细胞对Ⅳ型胶原的黏附特性，评价重复利用Ⅳ型胶原以差速黏附法在分选表皮干细胞中的效果。 方法：实验于2004—09／2005-06在解放军总医院第一附属医院全军烧伤研究所基础部完成。①包皮来自解放军空军总医院泌尿外科收治的7-25岁行包皮环切术患者。②未包被胶原的培养瓶作为对照1组，首次Ⅳ型胶原和3次Ⅳ型胶原包被的培养瓶分别作为对照2组和实验组1，将包被Ⅳ型胶原并曾黏附细胞的培养瓶消化后作为实验组2。③人角质形成细胞分别接种其上，15min后对贴壁细胞进行显微摄像并计数；对贴壁细胞的生长及增殖活性进行观察；并用β1整合素、鼠抗人角蛋白19对贴壁的细胞进行鉴定。 结果：①胶原包被组的贴壁细胞数明显增多沪≤0．05），并于5d铺满培养瓶底，而对照1组2d后仍未铺满瓶底。对照1组15min细胞贴壁数显著低于对照2组、实验1组、实验2组[（119．0±3．0），（246．3±19．6），（241．3±15．3），（262．0±11．1）个／100倍视野，P≤0．01]；对照2组与各实验组差异无显著性意义（P〉0．05）。②贴壁细胞β1整合素和鼠抗人角蛋白19表达阳性。 结论：表皮干细胞对Ⅳ型胶原具有较好的黏附特性，回收利用Ⅳ型胶原对表皮干细胞的黏附特性几乎无影响，并可节约费用。

英文摘要：

AIM： To observe the characteristics of epidermal stem cells adhering to collagen Ⅳ and evaluate the effect of differential attachment to recycled collagen Ⅳ in isolation of epidermal stem cells. METHODS： This experiment was conducted at the Basic Department, General Institute of Bums, First Hospital Affiliated to General Hospital of Chinese PLA from September 2004 to June 2005. ① Foreskins were from the 7 to 25-year-old patients who undergone posthectomy in the Department of Urinary Surgery, Air Force General Hospital of Chinese PLA. ② Flasks coated nothing as control group Ⅰ ; Flasks coated type Ⅳ collagen which was firstly and thirdly used as control group Ⅱ and experimental group Ⅰ , respectively; Flasks that had coated type Ⅳ collagen and ever seeded cells were recycled after digested as experimental group Ⅱ. ③Keratinocytes were isolated from human circumcised foreskins and seeded in the flasks. 15 minutes later, the cells adhered were photomicrographed and counted. Then growth and proliferation of the cells were observed from the micrograph. β1 integrin and keratin 19 （mouse-antikeratin 19）, markers of epidermal stem cells, were employed to identify the adhering ceils. RESULTS： ① The number of adhered cells was significantly increased in the flasks coated type Ⅳ collagen （P ≤ 0.05）, and cells covered the bottom of the flasks fully on day 5 , but cells did not cover the bottom of the flasks fully 2 days later in the control groupⅠ . 15 minutes later , the adhered cells was significantly lower in control groupⅠ than in control group Ⅱ, experiment groupⅠ or experiment group Ⅱ [（119.0±3.0）,（246.3 ±19.6）,（241.3±15.3）,（262.0±11.1）/100 times field,P≤ 0.011 . There was no significant difference between control group Ⅱ and each experimental groups （P 〉 0.05）. ② The adhered cells express β1 intega-in and keratin 19 positive. CONCLUSION： Epidermal stem cells have good adhering characteristics to type Ⅳ collagen. Recycled co