Proliferative effect and osteoinductive potential of extracellular matrix coated on cell culture plates.
Proliferative effect and osteoinductive potential of extracellular matrix coated on cell culture plates.
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细胞培养板上包被的细胞外基质的增殖作用和骨诱导潜力
DOI：
10.1186/2193-1801-2-303
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Huang S
中科院分区：
文献类型：
--
作者：
Guo Y;Zeng Q;Yan Y;Shen L;Liu L;Li R;Zhang X;Wu J;Guan J;Huang S
不同细胞/组织来源的细胞外基质(ECM)表现出细微的差异，这可能为体外或体内细胞的增殖和分化提供重要线索。然而，不同的细胞外基质在体外的生物活性还不完全清楚。本研究通过对成骨细胞MC3T3-E1和大鼠成纤维细胞进行培养、脱细胞处理，分别制备了成骨细胞来源的细胞外基质和成纤维细胞来源的ECM包被细胞培养皿。我们用细胞、生化和分子方法研究了两种不同的细胞外基质在细胞培养板上的生物活性。骨髓间充质干细胞在成骨细胞-ECM上的增殖活性低于在成纤维细胞-ECM上生长的BMSCs。与成纤维细胞来源的ECM培养的BMSCs相比，成骨细胞ECM培养的细胞碱性磷酸酶(ALP)活性增强，BMP-2和骨桥蛋白水平升高，分泌的钙含量增加，矮小相关转录因子2(RUNX 2)和骨钙素(OCN)mRNA水平升高。通过shRNA基因表达下调BMP-2或FGF-2基因，对种植于成骨细胞-ECM或成纤维细胞-ECM上的MC3T3-E1成骨细胞的分化和增殖无明显影响。因此，成骨细胞ECM比成纤维细胞ECM具有更好的成骨诱导能力和更低的增殖能力，两种ECM具有足够的生物活性，抑制生长因子对再植细胞的分化和增殖无明显影响。
Different cell/tissue derived extracellular matrix (ECM) display subtle differences that might provide important cues for proliferation and differentiation of cells in vitro or in vivo. However, the bioactivities of different ECMs in vitro were not fully understood. In this study, osteoblasts-derived and fibroblast-derived ECM-coated cell culture dishes were prepared respectively by culturing osteoblastic MC3T3-E1 cells and rat fibroblast then decellularizing the cultures. We investigated the bioactivities of the two different ECMs coated on cell culture plates using cellular, biochemical and molecular method. The proliferative activity of the bone marrow-derived mesenchymal stem cells (BMSCs) cultured on osteoblast-ECM was lower than for BMSCs grown on fibroblast-ECM. Compared with the BMSCs cultured on fibroblast-derived ECM, the cells grown on osteoblastic ECM showed enhanced alkaline phosphatase (ALP) activity, higher BMP-2 and osteopontin protein levels, increased secreted calcium content, and higher levels of runt-related transcriptional factor 2 (Runx 2) and osteocalcin (OCN) mRNA. Knockdown of BMP-2 or FGF-2 with shRNA transfection hardly effected osteoblastic differentiation or proliferation of MC3T3-E1 seeded on osteoblast-ECM or fibroblast-ECM. Therefore, the osteoblastic ECM had better osteoinductive potential and lower proliferative effect than fibroblastic ECM, and the two ECM presented enough bioactivity, knockdown of growth factors had no significant effect on differentiation and proliferation of re-seeded cells.