摘要
目的:观察人肾小管上皮细胞在壳聚糖/明胶三维支架培养系统中的生长特性,构建人肾小管上皮细胞三维培养生长模型。方法:将人肾近曲小管上皮细胞接种于壳聚糖/明胶三维支架中培养,电镜观察培养不同时间后的细胞形态。用Annexin V&PI双染检测三维培养的细胞凋亡率。结果:电镜观察显示人肾小管细胞在壳聚糖/明胶三维支架中紧贴支架生长。三维培养细胞早期自发凋亡细胞率为2.53%,与二维培养细胞的3.26%相比显著减少(P<0.05);三维培养和二维培养细胞晚期凋亡细胞率分别为1.81%和2.61%,但两者差异无统计学意义(P>0.05)。结论:结果说明壳聚糖/明胶三维支架能够保持人肾小管上皮细胞的细胞活力,降低细胞早期凋亡率是进行人肾小管上皮细胞体外培养的有效途径。
Objective： To investigate the growth characteristics of renal proximal tubule cells cultured in the three - dimensional chitosan/ gelatin scaffold. To establish a three - dimensional chitosan/gelatin scaffold for renal proximal tubule cells culture. Method： Renal proxi-mal tubule cells were cultured in the three - dimensional chitosan/gelatin scaffold. Scanning electron microscope was used for the observa-tion of cell morphology. The cells were stained by Annexin V＆PI and apoptosis ratios were detected by flowcytomerty. Result： Renal prox-imal tubule cells grew in monolayer in the two dimensional culture system and the adhesion and interconnection of cells in the scaffolds are formed after cells seeded. The proportions of early stage of apoptosis cells were significantly lower in the cells cultured in three - dimen-sional chitosan/gelatin scaffold （2. 53% ） than those in the two -dimensional culture system （3.26%） （P 〈 0. 05 ）. Cells cultured in the three-dimensional chitosan/gelatin scaffold displayed lower ratios of late stage of apoptosis （1.81%） compared with that cultured in mznolayer （2.61% ） , while there was no significant difference （P 〉 0.05 ）. Conclusion： Renal proximal tubule cells cultured in the three - dimensional chitosan/gelatin scaffold displayed significantly lower ratio of early stage of apoptosis cells. This cell culture system can be used as a biomaterial plants for the investigation of tubular ceils proliferation after injury.
出处
《生物技术》
CAS
CSCD
北大核心
2013年第5期78-82,共5页
Biotechnology
关键词
肾小管上皮细胞
三维培养
细胞凋亡
组织工程
Renal tubular epithelial cells
3D culture
Apoptosis
Tissue engineering
