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目的建立人肺精确薄片体外移植培养模型;并检测暴露于流感病毒下的人肺薄片里是否存在感染和病毒复制。方法参照改良琼脂糖充填肺组织的方法,以1.5%低熔低胶琼脂糖经充膨、钻取、切片及培养等步骤建立人肺精确薄片体外移植培养模型。选取不同时间点对移植培养的人肺薄片进行显微镜下组织形态观察;以乳酸脱氢酶释放毒性实验评价细胞毒性;以RT-PCR和免疫组织化学方法测定人肺薄片组织对暴露的流感病毒的反应。结果人肺精确薄片模型经长时间(>10d)的培养,维持正常的结构和功能,支持人肺薄片组织暴露于流感病毒后的感染和病毒复制。结论建立的人肺精确薄片体外移植模型可靠和稳定,可被用于流感病毒与人体肺器官感染的研究。
OBJECTIVE: To establish an in vitro model of human lung implants in vitro and to detect the presence of infection and virus replication in human lung slices exposed to influenza virus. Methods According to the method of modified agarose filling lung tissue, in vitro culture model of human lung implants was established by 1.5% low melt low gel agarose by means of inflation, drilling, slicing and culture. Tissue morphology of the transplanted human lung slices was observed at different time points; the cytotoxicity was evaluated by the lactate dehydrogenase release toxicity test; the effect of human lung tissue on the exposure of influenza virus reaction. Results The model of human lung implants was cultured for a long time (> 10d), maintaining the normal structure and function and supporting the infection and virus replication of human lung tissue after exposure to influenza virus. Conclusion The established in vitro model of human lung implants in vitro is reliable and stable and can be used in the study of influenza virus and human lung organ infection.