聚对苯二甲酸乙二酯三维支架及碱性成纤维生长因子和乳酸对成人骨髓间充质干细胞增殖的影响

Influence of poly(ethylene terephthalate) three-dimensional scaffold,basic fibroblast growth factor and lactic acid on proliferation of human bone marrow mesenchymal stem cells

背景:诸多实验聚焦在改变传统的细胞培养方式,希望利用三维支架材料能够获得更多细胞量。目的:观察人骨髓间充质干细胞在三维培养条件下的增殖特点。方法:检测不同质量浓度的碱性成纤维生长因子和乳酸对人骨髓间充质干细胞增殖的影响。在确定最佳条件基础上,进一步测定聚对苯二甲酸乙二酯三维支架上人骨髓间充质干细胞的增殖和代谢的特点。通过Celltracker Green染色,激光共聚焦显微镜观察人骨髓间充质干细胞在三维支架材料上的活性。利用扫描电镜观察人骨髓间充质干细胞在三维支架材料上的形态分布特点。结果与结论:当培养基中碱性成纤维生长因子质量浓度为5μg/L,乳酸质量浓度少于0.9g/L时,最有利于促进三维支架材料上人骨髓间充质干细胞的增殖。在此优化条件下,人骨髓间充质干细胞在实验组(含5μg/LbFGF)和对照组(不含bFGF)中的扩增倍数分别为15.3和9.6,比生长速率分别为0.136/d和0.113/d,实验组的细胞量比对照组提高了160%。激光共聚焦显微镜和扫描电镜检测结果表明,人骨髓间充质干细胞在三维支架材料上呈立体、多层生长,聚对苯二甲酸乙二酯三维支架的特有结构为细胞提供相对较大的生长空间。

BACKGROUND：Many studies focus on the change of conventional cell culture condition,a three-dimensional scaffold could be used to support high density culturing.OBJECTIVE：To observe the proliferation of human bone marrow mesenchymal stem cells （hMSCs） in three-dimensional culture condition.METHODS：The effect of basic fibroblast growth factor and lactic acid on the proliferation of hMSCs was measured.Based on the optimized condition,the growth and metabolism of hMSCs cultured in poly（ethylene terephthalate） were further investigated;Dyed by Cell tracker Green,the activity of hMSCs-poly（ethylene terephthalate） was imaged under a laser confocal microscope;the details of hMSCs-poly（ethylene terephthalate） were observed using scanning electron microscope.RESULTS AND CONCLUSION：A low-does basic fibroblast growth factor （5 μg/L） and lactic acid （ 0.9 g/L） in culture medium displayed a significant promotion of the proliferation of hMSCs.Under the optimized condition,for the experimental group （with 5 μg/L basic fibroblast growth factor） and control （without 5 μg/L basic fibroblast growth factor）,the expansion folds and specific growth rate were 15.29 and 0.136,9.66 and 0.113,respectively.The amount of cells in the experimental group was increased 160% than that in control group.Observation of confocal laser microscopy and scanning electron microscope illustrated that the morphology and attachment of cells on poly（ethylene terephthalate） were stereoscopic,multi-level distributed,the unique structure of poly（ethylene terephthalate） three-dimensional scaffold provides cells with a larger growing space.