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密度梯度离心法分离培养犬骨髓基质干细胞(BM-SCs);将向成骨细胞诱导后的BMSCs接种至温度反应性培养皿中,37℃、5%CO2饱和湿度培养,然后降温至20℃制备BMSCs细胞片层;制备犬脱钙骨基质(DBM)及富血小板血浆(PRP);将DBM/PRP/BMSCs细胞片层/BMSCs植入犬左侧背阔肌深面、右侧相应部位植入DBM/PRP/BMSCs,观察其成骨效果。结果:当温度降至20℃时,BMSCs从温度反应性培养皿上完全脱落,形成细胞片层,将其覆盖于DBM/PRP/BMSCs,其成骨效果优于不加细胞片层的组织工程骨。结论:细胞片层技术与传统的骨组织工程方法相结合可构建出较理想的功能性组织工程骨。
Cultured canine bone marrow stromal cells (BM-SCs) were isolated by density gradient centrifugation. Osteoblast-derived BMSCs were inoculated into temperature-responsive culture dishes and incubated at 37 ° C in 5% CO2 saturated humidity before cooling to 20 ° C BMSCs cell sheets were prepared; DBM and PRP were prepared; DBM / PRP / BMSCs cell sheets / BMSCs were implanted deep into the left latissimus dorsi of dogs and corresponding parts of the right side were implanted DBM / PRP / BMSCs to observe the osteogenic effect. Results: When the temperature dropped to 20 ℃, the BMSCs completely detached from the temperature-responsive culture dish and formed a cell sheet that covered the DBM / PRP / BMSCs. The osteogenic effect was better than that without the cell sheet bone. Conclusion: The combination of cell sheet technique and traditional bone tissue engineering method can be used to construct the ideal functional tissue engineering bone.