人真皮成纤维细胞诱导分化研究

Induced differentiation of human dermal fibroblast

目的体外分离培养人包皮成纤维细胞(HFF),探讨其成骨、成脂、成软骨分化潜能,为骨创伤或其他修复治疗提供新的种子细胞来源。方法采用机械法结合酶消化法分离、培养HFF。取第3代细胞用免疫细胞化学法检测其波形蛋白(Vimentin)和角蛋白19(CK19)的表达。通过向成骨、成脂、成软骨细胞诱导分化(诱导培养基根据说明书配制),鉴定其多向分化潜能。结果原代细胞24h内完全贴壁,细胞形态不规则,多为圆形或多角形,传代后细胞形态均一,基本为长梭形,鱼群样、漩涡样生长。免疫细胞化学结果显示该细胞表达Vimentin,不表达CK19,经诱导可以向成骨、成脂、软骨细胞方向分化。结论 HFF在特定条件能多向分化,其本身也参与骨折愈合的全过程,加之来源方便,增殖能力强,有望为骨折或较大骨缺损患者修复治疗及组织工程学提供理想种子细胞。

Objective To separate and culture human foreskin fibroblasts（HFF） in vitro and to investigate the differentiation potentials of osteogenesis,adipogenesis and chondrogenesis to provide the new source of seed cells for bone trauma or other repair treatment. Methods The methanical method combined with the enzyme digestion method was adopted to separate and culture HFF. The third generation cells were taken for detecting the expression of Vimentin and CK19 by the immunocytochemical method. By the induction differentiation to osteoblast,adipoblast and chondroblast（the induction medium was prepared according to the in- structions）, the multi-directional differentiation potentials were identified. Results The primary cells were completely adherent within 24 h, the cell shapes were irregular, most of cells were round or polygonal, after passage, the cellular shapes were uniform, u- sually long spindle-shape,grew in fish shoal-like and swirl-like. The immunocytochemical results showed that these cells expressed Vimentin and not expressed CK19. After being cultured in inducing differentiation media, the cells were differentiated into the direc tions of osteoblasts,adipoblasts and chondroblasts. Conclusion HFF can multi-directionally differentiate under certain condition, which itself also participates in the whole process of fracture healing,together with a convenient source and high proliferation ability, HFF is expected to provide the ideal seed cells for the repair treatment of fracture or larger bone defect and the tissue engineer ing.