Reorganization of basement membrane matrices by cellular traction promotes the formation of cellular networks in vitro.
Reorganization of basement membrane matrices by cellular traction promotes the formation of cellular networks in vitro.
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发表时间：
1992-05
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通讯作者：
R. Vernon;J. Angello;M. Iruela-Arispe;T. Lane;E. Sage
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作者：
R. Vernon;J. Angello;M. Iruela-Arispe;T. Lane;E. Sage
血管内皮细胞在胶质基膜基质层上培养，迅速组织成索状或管状结构的网络。虽然这种现象是体内血管生成的潜在模型，但我们质疑基底膜基质是否以特定的方式指导内皮细胞的分化。在这项研究中，我们研究了影响体外细胞网络形成的因素，试图确定这一过程的基本机制。我们发现内皮细胞、成纤维细胞、平滑肌细胞和小鼠间质细胞系TM3细胞在基底膜基质上形成网络的方式大致相同。光学和电子显微镜，结合延时视频显微镜，揭示了细胞组织在排列基底膜基质的棋盘状网络上，这是由细胞牵引力的张力产生的。细胞伸长和通过凝胶表面的渐进式运动被限制在排列的基质轨道上，直到轨道出现才发生。基底膜基质上细胞网络的形成可以通过降低基质厚度、在基质中加入原生I型胶原或破坏细胞骨架微丝和微管来抑制。细胞分裂不需要网络的形成。形成网络的牛主动脉内皮细胞不能同时转录I型胶原的mRNA， I型胶原是一种由内皮细胞合成的蛋白质，在体外自发形成管状。此外，网络形成细胞中纤维连接蛋白和SPARC（酸性且富含半胱氨酸的分泌蛋白）的mRNA水平与未形成网络的内皮细胞中的水平相似。内皮细胞和TM3细胞被涂在天然I型胶原蛋白的高延展性凝胶上，也形成了索，并将基质纤维排列成线性轨迹，类似于基底膜基质上产生的，尽管结构不那么明确。我们的观察结果表明，细胞外基质的机械化学特性能够将细胞牵引的力量转化为指导复杂细胞模式形成的模板。
Vascular endothelial cells that are cultured on layers of gelled basement membrane matrix organize rapidly into networks of cords or tubelike structures. Although this phenomenon is a potential model for angiogenesis in vivo, we questioned whether basement membrane matrix directs the differentiation of endothelial cells in a specific manner. In this study, we have examined factors that influence the formation of cellular networks in vitro in an attempt to define a basic mechanism for this process. We found that endothelial cells, fibroblasts, smooth muscle cells, and cells of the murine Leydig cell line TM3 formed networks on basement membrane matrix in much the same fashion. Light and electron microscopy, combined with time-lapse videomicroscopy, revealed that cells organized on a tesselated network of aligned basement membrane matrix that was generated by tension forces of cellular traction. Cellular elongation and progressive motility across the surface of the gel were restricted to tracks of aligned matrix and did not occur until the tracks appeared. The formation of cellular networks on basement membrane matrix was inhibited by reducing the thickness of the matrix, by including native type I collagen in the matrix, or by disrupting cytoskeletal microfilaments and microtubules. Cell division was not required for network formation. Bovine aortic endothelial cells that formed networks did not simultaneously transcribe mRNA for type I collagen, a protein synthesized by endothelial cells that form tubes spontaneously in vitro. Moreover, levels of mRNA for fibronectin and SPARC (Secreted Protein that is Acidic and Rich in Cysteine) in network-forming cells were similar to levels seen in endothelial cells that did not form networks. Endothelial cells and TM3 cells that were plated on highly malleable gels of native type I collagen also formed cords and aligned matrix fibers into linear tracks that resembled those generated on basement membrane matrix, although the structures were not as well-defined. Our observations suggest that the mechanochemical properties of extracellular matrices are able to translate the forces of cellular traction into templates that direct the formation of complex cellular patterns.