体外壳聚糖—明胶—生长因子缓释支架对骨髓间充质细胞增殖的影响

发布时间：2018-05-20 01:28

本文选题：骨髓间充质干细胞 + 生长因子　；参考：《南昌大学》2010年硕士论文

【摘要】： 目的：制备具有缓释功能的壳聚糖—明胶—碱性成纤维细胞生长因子(bFGF)—肝细胞生长因子(HGF)三维大孔支架,探讨两种生长因子在体外对骨髓间充质干细胞(BMSCs)增殖的协同作用。方法：采用冷冻干燥法,将不同比例的壳聚糖、明胶、bFGF和HGF依次混合,通过冷冻、干燥,使其成为具有一定孔径的三维缓释支架。取SD大鼠乳鼠股骨和胫骨骨髓,分离、培养骨髓间充质干细胞。取生长状态良好的第三代BMSCs,将其接种于96孔板后,加入支架混合培养,5d后进行MTT细胞增殖测定。实验过程中对动物的处理符合伦理学标准。结果：在与含1μg/mL的bFGF支架混合培养,以及与同时含1μg/mL的bFGF、1μg/mL的HGF支架混合培养后,BMSCs明显增殖(P0.05),但这二组间无差别(P0.05)。而与含0.1μg/mL的bFGF支架、0.01μg/mL的bFGF支架、1μg/mL的HGF支架混合后,细胞增殖无统计学差异(P0.05)。结论：梯度密度离心法结合贴壁筛选法能获得高纯度高活性BMSCs;壳聚糖—明胶可作为生长因子缓释支架材料；bFGF具有促进BMSCs增殖的作用,促进作用的大小与加入bFGF的量有关；HGF对BMSCs不具有增殖作用；bFGF和HGF在体外促进BMSCs增殖上不具有协同性。
[Abstract]:Aim: to study the synergistic effect of two kinds of growth factors on the proliferation of bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: by freeze-drying method, chitosan, gelatin bFGF and HGF were mixed in turn to form a three-dimensional sustained-release scaffold with a certain pore diameter by freeze-drying. Bone marrow of femur and tibia were isolated from neonatal SD rats and bone marrow mesenchymal stem cells were cultured. The third generation BMSCs with good growth state were inoculated on 96-well plate and mixed with scaffold for 5 days to determine the proliferation of MTT cells. The treatment of animals in the course of the experiment accords with ethical standards. Results: after mixed culture with bFGF scaffolds containing 1 渭 g/mL and HGF scaffolds containing 1 渭 g/mL and bFGF1 渭 g/mL, the proliferation of BMSCs was significantly increased (P 0.05), but there was no difference between the two groups (P 0.05). However, when mixed with 0.1 渭 g/mL bFGF scaffold or 0.01 渭 g/mL bFGF stent or 1 渭 g/mL HGF scaffold, there was no significant difference in cell proliferation (P 0.05). Conclusion: high purity and high activity BMSCs can be obtained by gradient density centrifugation combined with adherent screening. Chitosan gelatin can be used as a growth factor sustained-release scaffold material to promote the proliferation of BMSCs. The effect of bFGF was related to the addition of bFGF to BMSCs. BFGF and HGF had no synergistic effect on the proliferation of BMSCs in vitro.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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