Wohlwend A, Montesano R, Vassalli J D, Orci L
J Cell Physiol. 1985 Dec;125(3):533-9. doi: 10.1002/jcp.1041250325.
In the present work, we have taken advantage of the properties of two recently isolated clonal subpopulations of the pig kidney-derived LLC-PK1 cell line to study aspects of the establishment of epithelial polarity. When grown in suspension, LLC-PK1/D + Sc cells reaggregated within a few hours and, during the following days of culture, formed free-floating, hollow spheres or cysts, lined by a monolayer of polarized cells. In contrast, LLC-PK1/D- cells were unable to develop such polarized structures even upon prolonged culture in suspension. The polarity of the LLC-PK1/D + Sc cells lining the cysts was inverted compared to that in intact renal tubules, the microvilli-rich "apical" pole being oriented toward the external medium. However, upon embedding these preformed cysts in collagen gels, a reversal of polarity was observed within hours, the microvilli-rich pole now facing the cyst cavity. Thus, in the same clonally derived cell population, cell-to-cell contact and interaction with the extracellular matrix differentially affect the orientation of cellular polarity. The LLC-PK1/D + Sc cysts provide a suitable in vitro model system for further study of the sequential events by which extracellular matrix components induce an appropriately oriented polarization. In addition, the comparison between LLC-PK1/D + Sc and D- cells, which differ in their ability to polarize in response to cell-to-cell contact, should help define some of the cellular determinants involved in epithelial organization.
在本研究中,我们利用猪肾来源的LLC-PK1细胞系最近分离出的两个克隆亚群的特性,来研究上皮极性建立的相关方面。当在悬浮培养时,LLC-PK1/D + Sc细胞在数小时内重新聚集,并且在随后的培养天数中,形成自由漂浮的中空球体或囊肿,由单层极化细胞排列。相比之下,LLC-PK1/D-细胞即使在悬浮培养延长后也无法形成这种极化结构。与完整肾小管中的情况相比,囊肿内衬的LLC-PK1/D + Sc细胞的极性发生了反转,富含微绒毛的“顶端”极朝向外部培养基。然而,将这些预先形成的囊肿嵌入胶原凝胶中后,数小时内观察到极性反转,富含微绒毛的极现在面向囊肿腔。因此,在同一克隆来源的细胞群体中,细胞间接触和与细胞外基质的相互作用对细胞极性的取向有不同的影响。LLC-PK1/D + Sc囊肿为进一步研究细胞外基质成分诱导适当取向极化的一系列事件提供了合适的体外模型系统。此外,LLC-PK1/D + Sc细胞和D-细胞之间的比较,它们在响应细胞间接触而极化的能力上有所不同,应该有助于确定一些参与上皮组织形成的细胞决定因素。