Published: April 27th, 2021
Here we demonstrate a method to apply fluid shear stress to cancer cells in suspension to model the effects of hemodynamic stress on circulating tumor cells.
During metastasis, cancer cells from solid tissues, including epithelia, gain access to the lymphatic and hematogenous circulation where they are exposed to mechanical stress due to hemodynamic flow. One of these stresses that circulating tumor cells (CTCs) experience is fluid shear stress (FSS). While cancer cells may experience low levels of FSS within the tumor due to interstitial flow, CTCs are exposed, without extracellular matrix attachment, to much greater levels of FSS. Physiologically, FSS ranges over 3-4 orders of magnitude, with low levels present in lymphatics (<1 dyne/cm2) and the highest levels present briefly as cells pass through the heart and around heart valves (>500 dynes/cm2). There are a few in vitro models designed to model different ranges of physiological shear stress over various time frames. This paper describes a model to investigate the consequences of brief (millisecond) pulses of high-level FSS on cancer cell biology using a simple syringe and needle system.
Metastasis, or the spread of cancer beyond the initial tumor site, is a major factor underlying cancer mortality1. During metastasis, cancer cells utilize the circulatory system as a highway to disseminate to distant sites throughout the body2,3. While en route to these sites, circulating tumor cells (CTCs) exist within a dynamic fluid microenvironment unlike that of their original primary tumor3,4,5. It has been proposed that this fluid microenvironment is one of many barriers to meta....
1. Cell preparation
Elevated resistance to FSS-induced mechanical destruction has been previously shown to be a conserved phenotype across multiple cancer cell lines and cancer cells freshly isolated from tumors relative to non-transformed epithelial cell comparators15,24. Here, additional cancer cell lines from a variety of tissue origins (Table 2) were tested to demonstrate that the majority of these cells display viability ≥ 20% after 10 pulses of FSS at 25.......
This paper demonstrates the application of FSS to cancer cells in suspension using a syringe and needle. Using this model, cancer cells have been shown to be more resistant to brief pulses of high-level FSS relative to non-transformed epithelial cells15,22,24. Furthermore, exposure to FSS using this model results in a rapid increase in cell stiffness, activation of RhoA, and increased cortical F-actin and myosin II-based contrac.......
|14 mL round bottom tubes
|Falcon - Corning
|30 G 1/2" Needle
|5 mL syringe
|96-well black bottom plate
|Costar - Corning
|BSA, Fraction V
|Cell Titer Blue
|Sodium Azide (NaN3)
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