Computerized analysis of tumor cells flowing in a parallel plate chamber to determine their adhesion stabilization lag time

Abstract

The importance of cell adhesion in a variety of physiological phenomena requires development of an understanding of the factors and molecular mechanisms underlying these behaviors. Cell adhesion is a multistep process involving primary receptor-ligand interactions followed by secondary events that may lead to the formation of focal contacts. Due to the lack of well-defined assays to study adhesion stabilization, little is known about this process, except that it may involve signaling events, receptor recruitment, and, as we have demonstrated, covalent peptide cross-linking by cell membrane-associated transglutaminase [Menter et al.: Cell Biophys. 18:123-143, 1992). To study the stabilization process we have developed a dynamic assay employing a parallel plate flow chamber coupled with video microscopy and digital image processing. Our studies utilize wheat germ agglutinin-selected human metastatic melanoma cell variants that exhibit differences in their experimental metastatic potential and expression of transglutaminase. Using this assay, quantifying cell-substrate stabilization was found to be quick, reliable, reproducible, and useful in evaluating agents that block this process.

Role of β3 Integrins in Melanoma Cell Adhesion to Activated Platelets under Flow

“…␣5␤1 and ␣v integrins were shown to support tumor cell arrest under flow to fibronectin or vitronectin substrates (68). Stabilization of the initial binding events seemed to depend on the activity of transglutaminase expressed by the tumor cells (69). Under our experimental conditions, one likely candidate to contribute to melanoma cell interaction with thrombi is P-selectin.…”

Effect of adsorbed von Willebrand factor and fibrinogen on platelet interactions with synthetic materials under flow conditions

“…Two types of flow cells were used in this work. For the PET and tetraglyme‐coated PET studies, a Sakariassen20, 21 flow cell system was used, while for the studies with polystyrene and glass, a GlycoTech flow cell22–25 was used. The motivation of using two flow cells was practical: the Glycotech cell is simply better, in using less blood, and ease of use (see Discussion), but did not become available to us until later in our work.…”

Src protein kinase pp60c‐src influences adhesion stabilization of HT‐29 colon carcinoma cells to extracellular matrix components under dynamic conditions of laminar flow

“…Flow rates were decreased in 30‐s intervals so that the WSS equaled 4.8, 4.0, 3.2, or 2.4 dynes/cm 2 . The WSS at the point of initial cell adhesion was used to measure wall shear adhesion threshold (WSAT) (29). The dynamic adhesion rate (DAR) represented the total number of adherent cells after 1 min at a low flow rate of 2.4 dynes/cm 2 , which was approximately 50% of WSAT and in the physiological range found in the microcirculation (1.5–3.0 dynes/cm 2 ).…”