体外培养所形成的细胞外基质对MC3T3-E1细胞生长和分化的影响

Effects of extracellular matrix produced in vitro on growth and differentiation of MC3T3-E1 cells

细胞外基对组织细胞起支持、保护、营养作用,对细胞的增殖、分化有重要影响,在细胞和组织工程中,应该充分考虑细胞外基质的作用。本研究首先脱去培养板中融合培养的原代小鼠心肌成纤维细胞和成骨细胞,获得两种体外形成的细胞外基质包被的培养板,其中成骨细胞细胞外基质中含有骨形成蛋白2。然后将MC3T3-E1成骨前体细胞接种在这种培养板中,发现成纤维细胞胞外基质包被的培养板中的细胞增殖活性最高,而成骨细胞胞外基质包被的培养板中细胞的碱性磷酸酶活性、骨形成蛋白2和骨桥蛋白的相对蛋白表达量最高,细胞外钙沉积量比其他组高1倍左右。结果表明:包被在培养板上的这两种细胞外基质有不同的生物活性,成纤维细胞胞外基质可促进成骨前体细胞增殖,成骨细胞胞外基质可促进成骨前体细胞骨向分化。

Extracellular matrix（ECM） keeps cell＇s shape,protects and nourishes cells;it plays a great role in cell proliferation and differentiation.Therefore,ECM is very important in cell and tissue engineering.In this study,after primary mouse osteoblasts and fibroblasts maintained at confluence in vitro were removed,their ECM coated on cell culture plate was prepared,and bone morphogenetic proteins 2（BMP-2） was detected in the osteoblasts ECM.MC3T3-E1 preosteoblasts cells were seeded on cell culture plates covered with fibroblasts ECM and osteoblasts ECM respectively.The proliferative activity of the cells cultured on fibroblasts ECM was higher than that on osteoblasts ECM and the control group.The alkaline phosphatase activity,relative protein levels of BMP-2 and osteopontin,secreted calcium of the cells cultured on osteoblasts ECM were all the highest.The results indicate that the two different ECMs produced in vitro had different bioactivities,the fibroblasts ECM coated on cell culture plates could accelerate MC3T3-E1 cells proliferation,and the osteoblasts ECM could promote cells osteogenic differentiation.