目的比较人循环纤维细胞与人胚肺成纤维细胞的形态学特征和合成胶原的能力。方法体外分离、培养人循环纤维细胞,用酶水解法和天狼星红染色法分别测定培养液中的羟脯氨酸含量和细胞层中的胶原含量,比较其在不同分化阶段与培养的人胚肺成纤维细胞合成胶原的能力,同时用Western blotting分析其分泌胶原的类型。结果循环纤维细胞在体外分化过程中,形态学特征不断变化,从圆形幼小、多聚集成“细胞岛”的纤维细胞向梭形、透明、边缘多纤毛状突起的成纤维细胞样细胞分化,在分化过程中不断合成胶原,尤其在培养的细胞层中伴有大量胶原沉积,所分泌的胶原以Ⅰ型和Ⅲ型胶原为主。结论循环纤维细胞能够在体外分化为成纤维细胞样细胞,与人胚肺成纤维细胞具有相似的胶原合成能力。
Objective To compare the morphological characters and collagen synthesis between circulating fibrocytes and fibroblasts. Methods Circulating fibrocytes were isolated from human peripheral blood by centrifugation over Ficoll-Paque and cultured in vitro. Then the collagen secreted into the medium was detected by the measurement of the hydroxyproline concentration with enzymolysis, and the collagen accumulated on the cell layer was determined by the optical density at 540nm of picro-sirius red staining. The contents of collagen were assessed by Western blotting. Results The morphological characteristics of circulating fibrocytes were varying when cultured in vitro and the collagen types Ⅰand Ⅲ were synthesized and secreted into the medium. Conclusion Circulating fibrocytes can differentiate into fibroblast-like cells and have the same ability of collagen synthesis compared with pulmonary fibroblasts of human fetus.