Collagen type Ⅲ is an important linking molecule between generated renal tubules and an artificial polyester interstitium--Reticulin as a linker between generated tubules and the interstitium  

作　　者：W.W. Minuth L. Denk A. Blattmann Ho Castrop 

机构地区：[1]Department of Molecular and Cellular Anatomy,University of Regensburg,University Strect 31,D-93053 Regensburg,Germany [2]Department of Physiology, University of Regensburg,University Strect 31,D-93053 Regensburg,Germany

出　　处：《中国组织工程研究与临床康复》2008年第32期6201-6208,共8页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：在再生医学领域,已经有很多人考虑在将来用干/祖细胞治疗急慢性肾功能衰竭。但是,要使这种治疗方案行之有效,需要了解关于患病肾脏肾小管发育的细胞生物学信息。与干/祖细胞治疗急慢性肾功能衰竭相关尚待明晰的细胞生物学问题包括:①干/祖细胞的整合。②干/祖细胞的定向分化类型。③新形成肾小管的空间构成。为了更好的了解这项技术的相关机制,文章应用了先进的培养技术在人工非细胞外基质材料界面条件下构建肾小管。将新生兔肾来源的干/祖细胞用多层聚酯纤维网覆盖,放置在灌注培养器中,用含1×10-7mol/L醛固酮的新鲜合成的在DMEM基础上改良的IMDM培养基诱导培养13d。扫描电镜下,在大豆凝集素、银染和抗Ⅲ型胶原或层粘连蛋白γ1单克隆抗体标记的全标本包埋爬片或冰冻切片上观察肾小管的生长情况。扫描电镜观察结果显示,新生成的肾小管完全被基底膜覆盖,纤维网状板有大量的纤维与生成的肾小管基底面和周围人工聚酯材料相互交联。抗Ⅲ型胶原染色和银染结果表明,在新生成的肾小管的基底膜和邻近的人工聚酯材料间有大量的纤维交织,在发育成熟的肾小管中有与新生成肾小管相同排列形式的Ⅲ型胶原纤维。以上实验结果说明,Ⅲ胶原是联系新生成的肾小管基底面和人工基质聚酯纤维的分子纽带。In the field of regenerative medicine, much consideration has been given to stem/progenitor cells for the future treatment of acute and chronic renal failure. For this strategy to be effective, however, cell biological information about tubule development within the diseased organ is needed. Unresolved cell-biological issues relating to this kind of treatment include （1） the integration of stem/progenitor cells, （2） their differentiation into site-specific cell types, and （3） the spatial formation of new tubules. To better understand the mechanisms related to this technology, renal tubules were generated at the interphase of an artificial interstitium by using advanced culture techniques. Stern/progenitor cells derived from neonatal rabbit kidney were covered with layers of polyester fleece, placed in a perfusion culture container, and superfused for 13 days with flesh and chemically defined Iscove＇s Modified Dulbeccos Medium （IMDM） containing aldosterone （1 × 10^-7 mol/L）. The spatial growth of tubules was registered by scanning electron microscopy （SEM） and on whole mounts or cryosections labeled with soybean agglutinin, silver stain and monoclonal antibodies reacting with collagen type Ⅲ or laminin γ1. SEM revealed that the generated tubules were completely covered by a basal lamina. The lamina fibroreticularis exhibited numerous fibers connecting the basal aspect of generated tubules with the surrounding polyester fibers. Cryosections labeled with monoclonal antibodies anti-collagen type Ⅲ and silver stain demonstrated the formation of numerous fibers spanning between the basal lamina of generated tubules and neighboring polyester fibers. In matured kidney tubules the same arrangement of collagen type Ⅲ fibers is observed as that for generated tubules. This work shows that collagen type Ⅲ is a relevant molecular linker between the basal aspect of generated renal tubules and the polyester fibers of the artificial interstitium.

关 键 词：组织工程学 灌注培养 干/祖细胞 肾 小管 人工间质 基底膜 Ⅲ型胶原 聚酯 

分 类 号：R318[医药卫生—生物医学工程]