小鼠胰岛与肝脏脱细胞支架三维共培养的实验研究

发布时间：2019-03-17 11:25

【摘要】：目的 :观测原代小鼠胰岛在小鼠肝脏脱细胞支架上的生长及功能,探讨组织工程学治疗糖尿病的新方法。方法 :运用灌注法制备结构完整的全肝脏脱细胞支架并检测,运用胶原酶P灌注消化法获取小鼠胰岛细胞并行结构和功能完整性分析;将分离纯化的原代小鼠胰岛经门静脉灌入全肝脏脱细胞支架,在三维培养体系培养后进行相关检测。结果:肝脏脱细胞支架检测未见细胞结构,胶原结构保存完整。支架内剩余DNA定量检测结果为(38±11)ng/mg ds DNA,生物相容性良好。双硫腙(dithizone,DTZ)染色胰岛呈现特异性猩红色,葡萄糖刺激实验提示在高糖组中培养的原代小鼠胰岛胰岛素分泌功能明显高于低糖组,差异具有统计学意义(P0.01);将胰岛移植入肝脏脱细胞支架培养5 d后胰岛定植,且胰岛素基因表达显示脱细胞支架三维培养中胰岛素表达水平与平面培养相比,差异具有统计学意义(P0.01)。结论:肝脏脱细胞支架三维培养体系中的胰岛细胞比传统平面培养具有更佳的细胞活力及更有效的胰岛素分泌功能。
[Abstract]:Aim: to observe the growth and function of primary mouse islets on mouse liver acellular scaffolds and to explore a new method of tissue engineering in the treatment of diabetes mellitus. Methods: the whole liver acellular scaffolds with intact structure were prepared by perfusion and detected. The pancreatic islet cells were obtained by collagenase P perfusion digestion method and the structure and functional integrity of pancreatic islet cells were analyzed. The isolated and purified primary mouse islets were infused into the whole liver acellular scaffolds through portal vein, and the correlation was detected after culture in three-dimensional culture system. Results: no cellular structure was found in the liver acellular scaffolds, and the collagen structure was preserved intact. The biocompatibility of (38 卤11) ng/mg ds DNA, was better than that of in-stent residual DNA. Pancreatic islets stained with dithizone (dithizone,DTZ) showed specific scarlet color. Glucose stimulation test showed that the insulin secretion function of primary mice cultured in high glucose group was significantly higher than that in low glucose group (P0.01). Pancreatic islets were transplanted into liver acellular scaffolds for 5 days, and insulin gene expression showed that insulin expression level in three-dimensional culture of acellular scaffolds was significantly higher than that in horizontal culture (P0.01). Conclusion: pancreatic islet cells in the three-dimensional culture system of liver acellular scaffolds have better cell viability and more effective insulin secretion function than those in the traditional planar culture system.
【作者单位】：南通大学附属医院普外科;苏州大学附属第一医院普外科;
【基金】：国家自然科学基金资助(81471801)
【分类号】：R318.08

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