Munn L L, Melder R J, Jain R K
Department of Radiation Oncology, Harvard Medical School, Boston, Massachusetts.
Biophys J. 1994 Aug;67(2):889-95. doi: 10.1016/S0006-3495(94)80550-8.
The parallel plate flow chamber provides a controlled environment for determinations of the shear stress at which cells in suspension can bind to endothelial cell monolayers. By decreasing the flow rate of cell-containing media over the monolayer and assessing the number of cells bound at each wall shear stress, the relationship between shear force and binding efficiency can be determined. The rate of binding should depend on the delivery of cells to the surface as well as the intrinsic cell-surface interactions; thus, only if the cell flux to the surface is known can the resulting binding curves be interpreted correctly. We present the development and validation of a mathematical model based on the sedimentation rate and velocity profile in the chamber for the delivery of cells from a flowing suspension to the chamber surface. Our results show that the flux depends on the bulk cell concentration, the distance from the entrance point, and the flow rate of the cell-containing medium. The model was then used in a normalization procedure for experiments in which T cells attach to TNF-alpha-stimulated HUVEC monolayers, showing that a threshold for adhesion occurs at a shear stress of about 3 dyn/cm2.
平行板流动腔为测定悬浮细胞与内皮细胞单层结合时的剪切应力提供了一个可控环境。通过降低含细胞培养基在单层上的流速,并评估在每个壁面剪切应力下结合的细胞数量,可以确定剪切力与结合效率之间的关系。结合速率应取决于细胞向表面的输送以及细胞与表面的固有相互作用;因此,只有在知道细胞向表面的通量时,才能正确解释所得的结合曲线。我们基于腔室内的沉降速率和速度分布,提出并验证了一个数学模型,用于将流动悬浮液中的细胞输送到腔室表面。我们的结果表明,通量取决于总体细胞浓度、距入口点的距离以及含细胞培养基的流速。然后,该模型被用于对T细胞附着于TNF-α刺激的人脐静脉内皮细胞(HUVEC)单层的实验进行归一化处理,结果表明在约3达因/平方厘米的剪切应力下出现了粘附阈值。