人增生性瘢痕皮肤与正常皮肤真皮间充质干细胞生物学特性的比较研究  

Comparative study of biological characteristics of dermis mesenchymal stem cells from human hy- pertrophic scar skin and normal skin

作　　者：周树萍[1] 徐静静[2] 蔡景龙 牛扶幼[1] 宗宪磊 都乐 

机构地区：[1]郑州大学第一附属医院整形美容外科,450052 [2]青海大学医学院附属医院烧伤整形科 [3]中国医学科学院整形外科医院瘢痕综合治疗中心

出　　处：《中华创伤杂志》2014年第5期388-393,共6页Chinese Journal of Trauma

基　　金：北京市自然科学基金资助项目（7122138）;中国医学科学院整形医院院所基金重大资助项目（5201010402）

摘　　要：目的比较人成熟期增生性瘢痕（hypertrophic scar，HS）皮肤和正常皮肤中真皮问充质干细胞（dermis mesenchymal stem cells，DMSCs）的生物学特性差异，为HS来源的DMSCs的临床应用及组织工程学研究奠定基础。方法取成熟期HS（瘢痕组）和正常皮肤（对照组）组织各20例，采用两步酶消化法分别提取和分选DMSCs，待两组细胞传代培养第3代时，分别观察其形态特点和检测其生长曲线；采用免疫细胞化学技术分别检测其细胞表面蛋白CD29、CD49及波形蛋白的表达情况；采用流式细胞术分别检测其细胞表面蛋白CD34、CD73、CD90及CD105表达阳性的细胞比例；采用RT—PCR法分别检测其Oct4基因和Nanog基因的表达，并利用诱导分化培养基分别检测其向成脂细胞、成骨细胞、成软骨细胞的分化能力。结果瘢痕组和对照组DMSCs形态和生长曲线基本一致，细胞标记物CD29、CD49、波形蛋白均呈阳性表达。瘢痕组和对照组CD34、CD73、CD90、CDl05表达分别为[（0．60±0．03）％：（0．61±0．02）％]、[（98．90±0．80）％：（99．00±0．70）％]、[（98．30±0．30）％：（98．20±0．40）％]、[（93．10±0．40）％：（93．00±0．20）％]（P均〉0．05）；Oct4、Nanog基因表达量分别为[（0．506±0．024）：（0．512±0．024）]和[（0．496±0．018）：（0．494±0．023）]（P均〉0．05）。体外诱导培养DMSCs均可向成脂肪细胞、成骨细胞、成软骨细胞分化。结论成熟期HS含有DMSCs，其生物学特性与正常皮肤DMSCs的生物学特性基本一致。Objective To lay a foundation for the clinical application and tissue engineering re- search of hypertrophic scar （HS） -derived DMSCs by comparing the biological characteristics of dermis mesenchymal stem cells （DMSCs） from human maturing-phase HS and normal skin. Methods Twenty maturing-phase HS specimens （scar group） and 20 normal skin specimens （control group） were selected to extract and sort DMSCs by two-step enzyme digestion. When cells in both groups were subcultured to 3rd generation, cell morphology and growth curve were observed; expressions of cell surface proteins CD29, CIM9 and vimentin were tested by immunocytochemistry; cells with positively expressed surface proteins CD34, CD73, CD90, and CD105 were examined by flow cytometry; expressions of genes Oct4 and Nanog were tested by RT-PCR; cell potential to differentiate into lipoblasts, osteoblasts, and chondroblasts was assayed in inductive medium. Results DMSCs in both groups showed similar shape and growth curve. Cell markers CD29, CD49 and vimentin expressed positively. Of scar and control groups, expressions of CD34, CD73, CD90, and CD105 were （0.60 ± 0.03） % vs （0.61 ± 0.02） %, （98.90±0.80）%vs （99.00±0.70）%, （98.30±0.30）%vs （98.20±0.40）%, and （93. 10± 0.40） % vs （ 93.00 ± 0.20） % respectively （ P 〉 0.05 ） ; expressions of genes Oct4 and Nanog were 0.506±0.024 vs 0.512±0.024 and0.496±0.018 vs 0.494±0.023 （P〉0.05）. Both types of DMSCs were able to differentiate in vitro into lipoblasts, osteoblasts, and chondroblasts in invitro conductive medium. Conclusion DMSCs exist in maturing-phase HS and present biomechanical characteristics basically similar with those of normal human skin.

关 键 词：真皮 瘢痕 间充质干细胞 

分 类 号：R622[医药卫生—整形外科]