Objective： To investigate the expression of growth differentiation factor 5（GDF-5） during limb skeletal development of mice and the effect of GDF-5 on bone marrow mesenchymal stem cells in vitro. Methods ： The exp...Objective： To investigate the expression of growth differentiation factor 5（GDF-5） during limb skeletal development of mice and the effect of GDF-5 on bone marrow mesenchymal stem cells in vitro. Methods ： The expression of GDF- 5 mRNA and protein in mouse fetal limb buds were detected in embryonic day 11.5-15.5 （E11.5-15.5） by RT-PCR and Western blotting respectively. Type Ⅱ collagen protein was examined with immunocytochemistry and the sulfate glycosaminoglycan was measured by Alcian blue. Results： During early stage of developmental skeletogenesis, the expression of GDF-5mRNA was constant and began with embryos E11.5, highlighted at embryos E12.5 and E13.5, subsequently dropped at embryos E14.5 and E15.5. There was very significant difference （P 〈 0.01） in average light density ratio of GDF-5/β-actin between E12.5-13.5 and the other three days. The expression of GDF-5 protein had a similar change with mRNA during limb skeletogenesis. Immunocytochemistry showed that GDF-5 could promote expression of Type Ⅱ collagen protein and histological staining of proteoglycan with Alcian blue revealed the deposition of typical cartilage extracellular matrix components. Conclusion： GDF-5 can enhance chondrogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro, which plays an important role in limb skeletal development and joint formation.展开更多
基金National Natural Science Foundation of China(No. 30471753)
文摘Objective： To investigate the expression of growth differentiation factor 5（GDF-5） during limb skeletal development of mice and the effect of GDF-5 on bone marrow mesenchymal stem cells in vitro. Methods ： The expression of GDF- 5 mRNA and protein in mouse fetal limb buds were detected in embryonic day 11.5-15.5 （E11.5-15.5） by RT-PCR and Western blotting respectively. Type Ⅱ collagen protein was examined with immunocytochemistry and the sulfate glycosaminoglycan was measured by Alcian blue. Results： During early stage of developmental skeletogenesis, the expression of GDF-5mRNA was constant and began with embryos E11.5, highlighted at embryos E12.5 and E13.5, subsequently dropped at embryos E14.5 and E15.5. There was very significant difference （P 〈 0.01） in average light density ratio of GDF-5/β-actin between E12.5-13.5 and the other three days. The expression of GDF-5 protein had a similar change with mRNA during limb skeletogenesis. Immunocytochemistry showed that GDF-5 could promote expression of Type Ⅱ collagen protein and histological staining of proteoglycan with Alcian blue revealed the deposition of typical cartilage extracellular matrix components. Conclusion： GDF-5 can enhance chondrogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro, which plays an important role in limb skeletal development and joint formation.
