细胞混合种植法构建生物人工肾小管的初步研究  

Mixing implantation for construction of bioartificial renal tubule:a preliminary study

作　　者：黄大伟[1] 傅博[1] 陈香美[1] 刘维萍[1] 汪杨[1] 

机构地区：[1]中国人民解放军总医院肾脏病研究所暨全军肾脏病重点实验室,北京100853

出　　处：《中国药物与临床》2008年第3期165-167,249,共4页Chinese Remedies & Clinics

基　　金：北京市科技计划基金资助项目(H060920051430)

摘　　要：目的为构建一种既有滤过及抗凝功能、同时又有重吸收及内分泌功能的新型生物人工肾小管提供实验基础。方法以层黏连蛋白0.74mg/ml包被的AV400滤器为载体,将转染人Nanog基因的血管内皮细胞(ECV304)悬液与转染人Nanog基因的肾小管上皮细胞(HKC)悬液等体积混匀,分次注入实验组滤器内腔,构建生物人工肾小管。对照组只在层黏连蛋白包被的AV400滤器内腔注入不含细胞的培养基。采用PKH26、PKH67标记法分别观察ECV304、HKC在聚砜膜中空纤维上的分布;应用扫描电镜检测混合细胞在聚砜膜中空纤维上的生长状况及形态。结果混合细胞能在聚砜膜中空纤维上较好地黏附、生长。PKH26、PKH67标记检测发现细胞呈致密点片状分布;扫描电镜观察可见细胞形成单层片状。结论两种转染细胞在层黏连蛋白包被的聚砜膜中空纤维上生长良好,这为构建一种既有血管内皮细胞抗凝、同时又有小管上皮细胞重吸收功能的新型生物人工肾小管奠定了实验基础。Objective To provide laboratory evidence for construction of a novel type of bioartificial renal tubule device which is potential with filtration, anticoagulation, re-absorption and endocrine activities. Methods Suspensions of hNanog gene transfected ECV304 cells and hNanog gene transfected HKC cells were prepared, then mixed in equal volumes and serially infused into the inner lumen （pre-coated with 0.74 mg/ml laminin） of an AV400 filter to construct the bioartificial renal tubule. As a control, cell-free culture medium was infused into another filter with the same setting described. Distribution of ECV304 cells and HKC cells on polysulfone hollow fibers retrieved were examined with PKH26/PKH67 cell labeling, and the growth and morphology of these cells was studied under a scanning electron microscope. Results Both transfected seeding cells were found with a satisfactory condition on the laminincoated polysulfone hollow fibers in terms of adherence and growth. PKH26/PKH67 labeling revealed a pattern of dense, patchy distribution, and scanning electron microscopy showed formation of a cell monolayer. Conclusion The favorable growth of transfected-ECV304 and transfected-HKC cells on laminin-coated polysulfone hollow fibers underlay a laboratory evidence for construction of a novel bioartificial renal tubule device which is potential with filtration, anticoagulation, re-absorption and endocrine activities.

关 键 词：转染 基因 Nanog 内皮细胞 上皮细胞 混合种植 聚砜膜中空纤维 肾小管 

分 类 号：R318.1[医药卫生—生物医学工程]