原位定量检测体外培养的尿道黏膜上皮细胞活性  

IN SITU QUANTITATION OF ACTIVITY OF CULTURED URETHRAL EPITHELIAL CELLS IN VITRO

作　　者：翟弘峰[1] 邱长虹[2] 张正文[1] 剑海燕[1] 康深松[1] 谢锋[1] 

机构地区：[1]河南省人民医院整形外科,郑州450003 [2]郑州市第一人民医院

出　　处：《中国修复重建外科杂志》2007年第12期1335-1337,共3页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：河南省杰出青年科学基金资助项目(0612000800);河南省卫生厅医学科技创新人才工程基金资助项目(2002203)~~

摘　　要：目的解决在不透明载体上观察细胞生长状况的难题。方法取雄性新西兰幼兔1只,进行尿道黏膜上皮细胞的原代培养并传至第3代。将其接种于胶原-壳聚糖复合材料上,体外培养3、7、14和21d后,行6-羧基二乙酸甲酯荧光素与碘化丙啶荧光双染,并应用激光共聚焦细胞仪检测。结果尿道黏膜上皮细胞在胶原-壳聚糖复合材料载体上生长,增殖良好。培养3、7d,活细胞荧光强度值为(1.09±0.13)×108、(2.04±0.13)×108,培养14、21d活细胞荧光强度值分别为(0.55±0.09)×108、(0.47±0.03)×108,与培养3、7d比较差异均有统计学意义(P<0.05)。各时间点死细胞荧光强度值差异无统计学意义(P>0.05)。结论利用激光共聚焦细胞仪对不同波长荧光强度分别进行定量分析,可以在原位定量快速检测尿道黏膜上皮细胞活性,动态观察组织工程组织生长情况,解决无法在不透明载体上观察细胞生长状况的难题。Objective To resolve the tough problem of bow to observe the growing cells in an opaque vector. Methods The urethral epithelial cells from a young male New Zealand rabbit were inoculated, and were primarily cultured in vitro and subcultured for 3 passages. Then, the urethral epithelial cells were cultured in the collagenchitosan complex for 3, 7, 14 and 21 days. The cells were dyed with 6-carboxyfluorescein diacetate-acetoxymethyl ester and propidium iodine, respectively. Then, Interactive Laser Cytometer was used to detect the growing cells. Results The urethral epithelial cells grew and proliferated very well in the collagen-ehitosan complex vector. After the urethral epithelial cells grew in the collagen-chitosan complex vector for 3 and 7 days, the fluorescent density amount of the surviving cells were （1.09±0.13）×10^8 and （2.04±0. 13）×10^8, respectively. However, after 14 and 21 days, the fluorescent density amount of the surviving cells was （0. 55 ±0.09） × 10^8 and （0. 47 ±0. 03） × 10^8, respectively. There was a significant difference when compared with the amount of the surviving cells at 3 and 7 days （P〈0.05）. Conclusion Using Interactive Laser Cytometer for measurement of the green and red fluorescent densities of different waves, the activity of the cultured urethral epithelial cells in vitro can be rapidly measured with the in situ quantitation method. This method solves a difficult problem of observing the growing cells in an opaque vector. The dynamic growing state of the engineering tissues can be observed.

关 键 词：激光共聚焦细胞仪 原位定量 尿道黏膜上皮细胞 

分 类 号：R699[医药卫生—泌尿科学]