山羊骨髓间充质干细胞向纤维软骨分化的形态学改变

Morphological changes of goat bone marrow mesenchymal stem cells differentiating into fibrochondrocytes

背景:前期初步研究发现,碱性成纤维细胞生长因子可诱导骨髓间充质干细胞向颞下颌关节盘细胞方向分化,且碱性成纤维细胞生长因子10μg/L诱导组合成胶原量明显高于5μg/L诱导组。目的:观察经不同浓度碱性成纤维细胞生长因子诱导后的骨髓间充质干细胞的超微结构变化。方法:原代分离培养山羊骨髓间充质干细胞,选P3,P4细胞,用5,10μg/L碱性成纤维细胞生长因子诱导骨髓间充质干细胞,以未加碱性成纤维细胞生长因子培养的骨髓间充质干细胞做对照。倒置相差显微镜观察细胞生长状况,用第7,14,21天的细胞爬片行蕃红O、天狼猩红和Ⅰ型胶原免疫组织化学染色,并观察第21天细胞的超微结构。结果与结论:经不同浓度碱性成纤维细胞生长因子诱导后,骨髓间充质干细胞可向颞下颌关节盘成纤维细胞样细胞形态分化,10μg/L组细胞更像关节盘成纤维细胞样细胞。提示骨髓间充质干细胞有向颞下颌关节盘细胞方向分化的形态学基础。

BACKGROUND：Our preliminary studies have shown that basic fibroblast growth factor can induce the differentiation of bone marrow mesenchymal stem cells into disc cells of the temporomandibular joint, and for basic fibroblast growth factor, 10μg/L is superior to 5μg/L in col agen synthesis. OBJECTIVE：To observe ultrastructural changes of bone marrow mesenchymal stem cells after being induced by different concentrations of basic fibroblast growth factor. METHODS：We cultured primary sheep bone marrow mesenchymal stem cells and selected passage 3 and 4 cells at good growth state. Bone marrow mesenchymal stem cells were stimulated with 5 and 10μg/L basic fibroblast growth factor and their growth state was observed under inverted phase contrast microscope. Uninduced cells served as controls. The slides with cellcrawling pieces were stained with Safranin O, picrosirius and type I col agen immunohistochemistry at days 7, 14 and 21, respectively. Simultaneously we col ected the cells at day 21 to observe the ultrastructural changes of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION：After being induced with different concentrations of basic fibroblast growth factor, bone marrow mesenchymal stem cells were able to differentiate into disc cells of the temporomandibular joint;and after being induced with 10μg/L basic fibroblast growth factor, cells were more＆amp;nbsp;like fibroblast-like cells of the temporomandibular joint disc. These findings indicate that bone marrow mesenchymal stem cells have morphological basis for differentiation to the fibroblast-like cells of the temporomandibular joint disc.