A fibrin or collagen gel assay for tissue cell chemotaxis: Assessment of fibroblast chemotaxis to GRGDSP
A fibrin or collagen gel assay for tissue cell chemotaxis: Assessment of fibroblast chemotaxis to GRGDSP
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DOI：
10.1006/excr.1998.4364
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发表时间：
1999-03-15
影响因子：
3.7
通讯作者：
Tranquillo, RT
中科院分区：
文献类型：
--
作者：
Knapp, DM;Helou, EF;Tranquillo, RT
成纤维细胞趋化性涉及许多生理过程，包括伤口愈合和形态发生。我们提出了一种新的测定成纤维细胞（和其他缓慢移动的组织细胞）的趋化性在一个直观的室含有生理相关的三维纤维蛋白或胶原蛋白凝胶，其中持久的，空间上连续的梯度已持续至少24小时，足够长的时间为显着的成纤维细胞迁移。这种特征组合在具有可比设置简单性的任何替代测定中不可用。使用假定的成纤维细胞趋化因子，纤连蛋白肽GRGDSP，我们测量了人包皮成纤维细胞在沿着梯度方向上的排列，其遵循对GRGDSP浓度的双相依赖性，最佳浓度约为10 nM。延时视频显微镜显示，细胞迁移的可溶性GRGDSP梯度，确认积极的趋化性GRGDSP和拒绝的可能性占主导地位的haptotaxis下降的可溶性GRGDSP梯度，也就是说，一个假定的梯度的整合素介导的粘附诱导的可溶性GRGDSP梯度。(C)北京：科学出版社.
Fibroblast chemotaxis is implicated in many physiological processes, including wound healing and morphogenesis. We present a novel assay for chemotaxis of fibroblasts (and other slow-moving tissue cells) in a direct-viewing chamber containing a physiologically relevant three-dimensional fibrin or collagen gel in which long-lasting, spatially continuous gradients have been sustained for at least 24 h, long enough for significant fibroblast migration. This combination of features is not available in any alternative assay of comparable setup simplicity. Using a putative fibroblast chemotactic factor, the fibronectin peptide GRGDSP, we measured human foreskin fibroblast alignment in the direction along the gradient, which followed a biphasic dependence on GRGDSP concentration with an optimal concentration of about 10 nM. Time-lapse video microscopy revealed that cell migration was up the soluble GRGDSP gradient, confirming positive chemotaxis to GRGDSP and rejecting the possibility of dominant haptotaxis down the soluble GRGDSP gradient, that is, up a putative gradient of integrin-mediated adhesion induced by the soluble GRGDSP gradient. (C) 1999 Academic Press.