文章摘要
王林林,杜为,张健,肖峰,汪道武,曹克将,王子盾,李小荣.优化贴壁法人皮肤成纤维细胞的原代培养[J].南京医科大学学报,2014,(1):
优化贴壁法人皮肤成纤维细胞的原代培养
Optimized Primary Culture of Human Skin Fibroblasts by Tissue Adherent Method
投稿时间:2013-06-18  修订日期:2013-11-04
DOI:10.7655/NYDXBNS201401017
中文关键词: 皮肤成纤维细胞 优化原代培养
英文关键词: human skin fibroblasts  optimized primary culture
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
作者单位E-mail
王林林 南京市胸科医院 linda81@139.com 
杜为 南京医科大学第一附属医院  
张健 南京医科大学第一附属医院  
肖峰 南京医科大学第一附属医院  
汪道武 南京医科大学第一附属医院  
曹克将 南京医科大学第一附属医院 KJcao@njmu.edu.cn 
王子盾 南京医科大学第一附属医院  
李小荣 南京医科大学第一附属医院  
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中文摘要:
      [摘要]目的:优化贴壁法原代培养人皮肤成纤维细胞的条件,建立稳定、经济、高效可行的人皮肤成纤维细胞培养方法。方法:采用包皮环切术后皮肤组织,剪成均匀组织块后贴壁,皮肤边缘长出团块状组织后胰酶消化、传代,并对所培养的细胞进行形态学观察及免疫荧光鉴定。此方法与酶消化法相对照。结果:优化贴壁法成功率100%,取得组织后2天皮肤边缘均可开始生长高密度细胞样物质,在一周内可获得高质量、高数量的细胞,传代贴壁后经镜下形态观察及细胞免疫荧光染色确定为皮肤成纤维细胞。结论:较之消化法,优化贴壁法细胞存活率高,活性及纯度好。该方法可稳定、经济地获得足够数量的细胞。
英文摘要:
      [ABSTRACT] Objective:To optimize and establish a stable, economical and feasible protocol for human skin fibroblasts primary culture. Methods:Foreskins were minced into pieces and placed into gelatin-coated 6-well plates. When there were lumpy tissues coming out, cells were digested and passaged. They were then observed by morphology and immunochemistry. This method was contrasted with enzyme digestion procedure. Results:The success rate of optimized tissue adherent method was 100%. Lumpy tissues consisted of High density cells grew from the edge of skin tissues within 2 days. High numbers of viable cells which were identified as fibroblasts by morphology and immunochemistry can be obtained within a week. Conclusion:Cells cultured by optimized tissue adherent method are with high survival rate, activity and purity. This method is more stable, economical and feasible in contrast with enzyme digestion procedure.
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