【摘要】 目的采用培养液循环式人工毛细管培养系统构建血管内皮细胞生物反应器,并对其形态和功能进行鉴定。方法将内皮细胞种植于无纺布中空纤维生物反应器的外腔,用培养液循环式人工毛细管培养系统进行培养,定期检测培养液中乳酸、葡萄糖、一氧化氮(nitrogen oxide,NO)和血管性血友病因子(von Willebr and Factor,vWf)的浓度。根据乳酸的浓度变化计算反应器内细胞的数量,通过葡萄糖的消耗和NO、vWf的产生反映内皮细胞的活力和功能。对其中的无纺布进行电镜、DAPI染色荧光显微镜检查,观察内皮细胞在生物反应器外腔无纺布上的生长情况。结果随着时间变化,培养液中的乳酸浓度进行性升高,升高的速度逐渐增大;葡萄糖浓度则逐渐降低,降低的速度逐渐增大;反映内皮细胞功能的NO在最初数小时轻度降低后也逐渐升高;在培养液中检测出内皮细胞的特异性标志物vWf;荧光显微镜和电镜下观察细胞在无纺布上生长良好,并具有内皮细胞的形态。结论血管内皮细胞能在无纺布中空纤维生物反应器外腔的无纺布上生长,并具有良好的形态和功能,提示未来在治疗脓毒症中的潜在价值。更多还原

【Abstract】 Objective To construct an endothelial bioreactor. Methods Endothelial cells were planted in the outer space of non-woven fabric polytetrafluoroethylene (PTFE) bioreactor with its intra-capillary space connecting to a medium perfusion system.The medium from the intra space was obtained every day and collected for lactic acid,glucose,nitrogen oxide (NO),protein C and von Willebrand Factor (vWF) analysis.Non-woven fabric PTFE was collected for fluorescent microscope and scanning electric microscope examination. Results Lactic acid levels in culture media sampling from the lumen space rose continuously and the rising rate was also increased.A contrary result was observed in glucose level.vWF can be detected in the extra space.The cells on the non-woven fabric PTFE had the character of endothelial cells under fluorescent microscope and scanning electric microscope examination. Conclusions Endothelial cells can be planted in the outer space of non-woven fabric PTFE hollow fiber bioreactor and have good shape and functions,suggesting its value in the therapy of sepsis.更多还原