摘要
目的构建大鼠脱细胞肾脏骨架,检测其内部细胞外基质成分的保留情况。方法取12周龄SD大鼠的肾脏,保留血管通路,从动脉依次灌入体积分数1%的十二烷基磺酸钠及体积分数1%的聚乙二醇辛基苯基醚溶液,脱细胞处理后利用HE染色及电镜观测其结构是否完整,是否有细胞残留,应用免疫组织化学染色检测脱细胞肾脏骨架细胞外基质成分保留是否完整。结果 HE染色显示脱细胞肾脏骨架内有完整的肾小球和肾小管结构,骨架内未见细胞成分残留;电镜结果显示脱细胞肾脏骨架中肾小球毛细血管基底膜连续完整,系膜基质得以保留;肾脏细胞外基质成分Ⅳ型胶原、层黏连蛋白、弹性蛋白的免疫组织化学染色显示出基本完整的肾脏骨架结构。结论向肾脏内灌注十二烷基磺酸钠及聚乙二醇辛基苯基醚溶液可成功构建出脱细胞的肾脏骨架,骨架内的肾小球和肾小管结构保留基本完整,无细胞成分残留,重要的细胞外基质成分也保留完整。
Objective To prepare rat decellularized kidney scaffolds and determine whether extracellular matrix ingredients are preserved. Methods Kidneys with vessels were harvested from 12-week old SD rats. 1% Sodium dodecyl sulfonate （SDS） and 1% Triton-X 100 were injected into kidneys from arteries successively. After the kidneys were decellularized, the scaffolds were observed under microscope with HE staining and transmitting electron microscope to determine whether the structure and extracellular matrix ingredients were intact. Results HE staining results indicated that gloumerular and tubular structure were remained intact and no ceil residue was found in the scaffolds. Transmission electron microscope identified that basement membrane was continual and intact, and mesangial matrix was also preserved. Immunohistochemistry staining results showed that scaffolds structure was stained intact by anti-type 1V, anti- elastin, and anti-laminin antibodies. Conclusions Decullularized kidney scaffold can be constructed by SDS and Triton-X 100 perfusion. No cell residue is found and the structure of scaffold is remained almost intact, and the important extracellular matrix ingredients are preserved.
出处
《中华实用诊断与治疗杂志》
2014年第5期447-449,共3页
Journal of Chinese Practical Diagnosis and Therapy
基金
海南省卫生厅科研课题重点项目(琼卫2013重点-15号)
国家自然科学基金面上项目(31170810
81102673)
北京科技新星计划(Z121107002512078)
关键词
脱细胞肾脏骨架
细胞外基质
组织工程
大鼠
Decellularized kidney scaffold
extracellular matrix
tissue engineering
rats
