摘要
目的探讨灌注法制备大鼠全肾脏脱细胞基质(ACM)支架的可行性。方法取12周龄的Wistar大鼠的肾脏,保留输尿管、肾静脉及肾动脉,沿肾动脉插入留置针建立灌注通道。灌注压强为100cmH2O,依次灌注肝素化PBS溶液,1%十二烷基磺酸钠(SDS)溶液,去离子水,1%TritonX-100溶液以及含青链霉素的PBS溶液。经脱细胞处理后,采用HE染色法光镜观察及扫描电镜观察支架微观结构,DAPI染色法荧光显微镜观察残留细胞核成分。结果所有经SDS及TritonX-100联合处理后的脱细胞基质支架在HE染色光镜观察下未发现细胞残留,扫描电镜观察仅见基膜及胶原蛋白等细胞外基质形成网状结构而无细胞,DAPI染色荧光显微镜观察未见DAPI与细胞核结合后所发出的强荧光。结论灌注法用于制备大鼠全肾脏脱细胞基质支架,简便可靠,该支架无细胞成分残留,是一种理想的细胞支架。
Objective To prepare rat whole-kidney acellular matrix （ACM） scaffolds using fluid perfusion method. Methods The kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH20. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining. Results No cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting ofbasilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope. Conclusion Fluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第5期979-982,共4页
Journal of Southern Medical University
关键词
灌注法
肾脏
脱细胞基质
组织工程
perfusion
kidney
acellular matrix
tissue engineering
