REPOPULATION OF A HUMAN ALVEOLAR MATRIX BY ADULT-RAT TYPE-II PNEUMOCYTES INVITRO - A NOVEL SYSTEM FOR TYPE-II PNEUMOCYTE CULTURE
REPOPULATION OF A HUMAN ALVEOLAR MATRIX BY ADULT-RAT TYPE-II PNEUMOCYTES INVITRO - A NOVEL SYSTEM FOR TYPE-II PNEUMOCYTE CULTURE
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DOI：
10.1016/0014-4827(86)90347-2
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发表时间：
1986-02-01
影响因子：
3.7
通讯作者：
MADRI, JA
中科院分区：
文献类型：
--
作者：
LWEBUGAMUKASA, JS;INGBAR, DH;MADRI, JA
本文介绍了肺脱细胞肺泡基质碎片的制备和大鼠II型肺细胞直接在肺泡上皮基底膜上的培养，从而可以研究肺基底膜对II型细胞形态和功能的影响。通过免疫荧光定位肺基质中的 I、III、IV 和 V 型胶原蛋白、层粘连蛋白和纤连蛋白，其模式与正常人肺的模式相同。碎片的透射电子显微镜（TEM）显示完整的上皮和内皮基底膜。通过阿尔新蓝染色，基质仍然存在。在 150 μm 厚的无细胞人肺泡细胞外基质碎片上培养的分离的成年大鼠 II 型肺细胞经历逐渐的细胞质扁平化，并丧失层状体、线粒体和表面微绒毛。这些变化类似于体内II型肺细胞分化为I型肺细胞。肺上皮基底膜上的 II 型肺细胞行为与人羊膜基底膜上培养的相同细胞类型的行为形成鲜明对比。在后一个表面上，细胞保持其立方体形状、层状体和表面微绒毛长达 8 天。这些观察结果表明，不同器官系统的基底膜对II型肺细胞的形态和功能产生不同的影响，并且肺泡和羊膜基底膜可能具有不同的三维组织。在肺基底膜上直接培养II型细胞的技术为研究基底膜对肺泡上皮细胞的调节作用提供了有用的工具。
This paper describes the preparation of lung acellular alveolar matrix fragments and culture of rat type II pneumocytes direct on on the alveolar epithelial basement membrane, thereby permitting study of the effect of lung basement membrane on the morphology and function of type II cells. Collagen types I, III, IV and V, laminin and fibronectin were located by immunofluorescence in the lung matrix with the same patterns as those described for the normal human lung. Transmission electron microscopy (TEM) of the fragments revealed intact epithelial and endothelial basement membranes. The matrix still present by Alcian Blue staining. Isolated adult rat type II pneumocytes cultured on 150 .mu.m thick fragments of acellular human alveolar extracellular matrix undergo gradual cytoplasmic flattening, with loss of lamellar bodies, mitochondria, and surface microvilli . These changes are similar to the in vivo differentiation of type II pneumocytes into type I pneumocytes. The type II pneumocyte behaviour on the lung epithelial basement membrane constrasted sharply with that of the same cell type cultured on a human amnionic basement membrane. On the latter surface the cells retained their cuboidal shape, lamellar bodies and surface microvilli for up to 8 days. These observations suggest that the basement membranes from different organ systems exert differing influences on the morphology and function of type II pneumocytes and that the alveolar and amnionic basement membranes may have differing three-dimensional organizations. The technique of direct culture of type II cells on the lung basement membrane provides a useful tool for studying the modulating effect of the basement membrane on alveolar epithelial cells.