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

doi:10.3969/j.issn.2095-4344.2014.06.007

Abstract

Abstract:

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, 10 μg/L is superior to 5 μg/L in collagen 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 10 μg/L basic fibroblast growth factor and their growth state was observed under inverted phase contrast microscope. Uninduced cells served as controls. The slides with cell crawling pieces were stained with Safranin O, picrosirius and type I collagen immunohistochemistry at days 7, 14 and 21, respectively. Simultaneously we collected 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 10 μg/L basic fibroblast growth factor, cells were more 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.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, mesenchymal stem cells, temporomandibular joint disk, fibroblast growth factor 2

CLC Number:

R394.2

Su Xue-lian, Bao Guang-jie, Kang Hong, Liu Lin, Kong Nan-nan. Morphological changes of goat bone marrow mesenchymal stem cells differentiating into fibrochondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(6): 860-865.

Figures/Tables 2

2.3 骨髓间充质干细胞诱导后组织学染色结果蕃红O染色：对照组细胞染色阴性，用固绿复染，以显示细胞轮廓，碱性成纤维细胞生长因子5 μg/L，10 μg/L组细胞染色均为阳性，细胞质着色呈紫红色，细胞核着色更深，说明经过碱性成纤维细胞生长因子诱导后的细胞有合成糖胺聚糖的能力(图2A，B)。天狼猩红染色：对照组细胞染色弱阳性，细胞质隐约可见，碱性成纤维细胞生长因子5 μg/L，10 μg/L组细胞染色均为强阳性，细胞质呈鲜红色，表明碱性成纤维细胞生长因子诱导后的细胞产生胶原的能力明显增强(图2C，D)。 Ⅰ型胶原免疫组织化学染色：对照组细胞染色阴性，苏木精复染细胞核呈蓝色，细胞质隐约可见；碱性成纤维细胞生长因子5 μg/L，10 μg/L组细胞染色均为阳性，细胞质和细胞核被染成棕黄色(图2E，F)。这表明经过碱性成纤维细胞生长因子诱导，细胞产生Ⅰ型胶原的能力逐渐增强，进一步证实诱导后的骨髓间充质干细胞合成和分泌Ⅰ型胶原的能力增强。 2.4 骨髓间充质干细胞超微结构观察结果对照组骨髓间充质干细胞透射电镜超微结构：电镜下骨髓间充质干细胞呈卵圆形，细胞核大、不规则，有指状突起，核仁多为1-3个，细胞质中有线粒体、粗面内质网和高尔基复合体，粗面内质网末端膨大成池，表面有核糖体(图3A，D)。碱性成纤维细胞生长因子 5 μg/L组透射电镜超微结构：骨髓间充质干细胞诱导21 d，细胞近似卵圆形，表面不规则，有多个指状突起，细胞核变小呈卵圆形，细胞质内细胞器更为丰富，线粒体和高尔基复合体明显增多，粗面内质网发达，还有大小不等的溶酶体，基质均匀，染色深浅不一，有的基质中可见更浓染的大颗粒，有的形成髓鞘样结构。在粗面内质网表面散在有大量的核糖体，有的核糖体聚集成池(图3B，E)。碱性成纤维细胞生长因子 10 μg/L组透射电镜超微结构：骨髓间充质干细胞诱导21 d，细胞近似圆形，突起少而短小，细胞核不规则，偏向一侧，细胞质内细胞器更为丰富，有大量的粗面内质网、高尔基复合体、线粒体、游离核糖体，有的核糖体聚集成池(图3C，F)。"

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