不同蛋白涂层在脱细胞牛心包片上对骨髓间充质细胞黏附生长的影响

为观察和评价不同蛋白涂层对骨髓间充质细胞在脱细胞牛心包片上黏附生长的影响,采用酶消化结合去污剂洗涤法制作脱细胞牛心包片,在其表面用不同蛋白(纤维连接蛋白、明胶蛋白、型胶原蛋白)均匀涂层。提取并培养大鼠骨髓间充质细胞,将其接种到预涂层的牛心包片上,未涂层组作为对照。在不同时间点采用Hochest染色观察细胞贴附生长情况,48h采用MTT法定量分析。纤维连接蛋白涂层及明胶蛋白涂层与型胶原蛋白涂层和不涂层组相比,对骨髓间充质细胞贴附增殖能力均有显著提高(P<0.001);纤维连接蛋白涂层和明胶蛋白涂层之间(P>0.05)以及型胶原蛋白涂层和不涂层对照组之间(P>0.05)对细胞的贴附生长没有显著差异。结果表明:脱细胞牛心包片上纤维连接蛋白涂层及明胶蛋白涂层能明显提高骨髓间充质细胞的贴附增殖能力,而型胶原蛋白涂层对骨髓间充质细胞的贴附增殖没有明显的改善。

Effect of Proteins Pre-coating on Bone Marrow Mesenchymal Cells Retention and Proliferation on Decellular Bovine Pericardium

To observe the growth of rat bone marrow mesenchymal cells (BMMCs) on decellular bovine pericardia in vitro and to investigate the effect of proteins pre-coating on cells retention and proliferation, bovine pericardia were decellularized using trypsin, DNase and Triton X-100 respectively. Then three proteins (fibronetin (FN), gelatin, collagen I) were coated on the surfaces of the bovine pericardia separately. BMMCs were harvested from rat thighbone marrow , then expanded and seeded onto decellular bovine pericardia with the proteins pre-coated . Decelluar bovine pericardia without coating were used as controls. The retention and growth of BMMSCs were observed by Hochest staining and analyzed by MTT method. It was shown that the retention and proliferation of BMMCs on FN group and gelatin group were significantly enhanced comparing with those on collagen I group and control group (P < 0.001). There was no significant difference between FN group and gelatin group (P > 0.05), nor between collagen I group and control group (P > 0.05). We conclude that the retention and proliferation of seeding cells on FN and gelatin could be significantly improved on decellular bovine pericardia (DBP) but not on collagen I.