摘要
目的研究软骨分化形成蛋白-2(CDMP-2)对小鼠骨髓基质干细胞软骨分化的作用。方法体外培养小鼠骨髓基质干细胞(MSCs),贴壁细胞传代,取第3代细胞,以不同浓度(0、10、20、50、100ng/ml)CDMP-2因子干预培养14d后,倒置相差显微镜观察细胞形态,RT-PCR、Westernblot和免疫细胞化学法检测不同浓度CDMP-2对Ⅱ型胶原表达的影响,阿尔辛蓝(Alcian)染色蛋白多糖。结果RT-PCR、Westernblot和免疫细胞化学显示经CDMP-2因子干预后,MSCs有Ⅱ型胶原mRNA和蛋白的表达,呈剂量依赖性;Alcian染色结果显示CDMP-2诱导细胞分泌蛋白多糖基质,高浓度组可见细胞小结区域呈明显的异染性。结论CDMP-2能够在体外定向诱导小鼠骨髓基质干细胞向软骨方向分化,50～100ng/ml为最佳剂量,为进一步探讨其在软骨发育机制中的作用提供了实验基础。
Objective To study the cartilage differentiation of mouse mesenchymal stem cells（MSCs） induced by cartilagederived morphogenetic proteins-2 （CDMP-2） in vitro. Methods The MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were chosen to induce into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2（0, 10, 20, 50 and 100 ng/ml）. After induction for 14 days, morphology of cells was observed under phase-contrast microscopy. Type Ⅱ collagen mRNA and protein were examined by using RT-PCR, Western blot and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue. Results CDMP-2 could promote the expression of Type Ⅱ collagen mRNA in a dose-dependant manner, especially strong with the concentrations of 50 and 100 ng/ml, and immunocytochemistry and Western blot revealed that the protein had a similar change. Histological staining proteoglycan of Alcian blue showed deposition of typical cartilage extracellular matrix. Conclusion Mouse bone marrow mesencymal stem cells can be differentiated into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the mechanism of CDMP-2 in chondrogenesis.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2008年第1期51-54,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No30471753)
关键词
软骨分化形成蛋白-2
骨髓基质干细胞
软骨分化
cartilage-derived morphogenetic proteins-2
bone marrow mesenchymal stem cells
chondrogenic differentiation
