The Viability of Single Cancer Cells after Exposure to Hydrodynamic Shear Stresses in a Spiral Microchannel: A Canine Cutaneous Mast Cell Tumor Model

Abstract

Our laboratory has the fundamental responsibility to study cancer stem cells (CSC) in various models of human and animal neoplasms. However, the major impediments that spike our accomplishment are the lack of universal biomarkers and cellular heterogeneity. To cope with these restrictions, we have tried to apply the concept of single cell analysis, which has hitherto been recommended throughout the world as an imperative solution pack for resolving such dilemmas. Accordingly, our first step was to utilize a predesigned spiral microchannel fabricated by our laboratory to perform size-based single cell separation using mast cell tumor (MCT) cells as a model. However, the impact of hydrodynamic shear stresses (HSS) on mechanical cell injury and viability in a spiral microchannel has not been fully investigated so far. Intuitively, our computational fluid dynamics (CFD) simulation has strongly revealed the formations of fluid shear stress (FSS) and extensional fluid stress (EFS) in the sorting system. The panel of biomedical assays has also disclosed cell degeneration and necrosis in the model. Therefore, we have herein reported the combinatorically detrimental effect of FSS and EFS on the viability of MCT cells after sorting in our spiral microchannel, with discussion on the possibly pathogenic mechanisms of HSS-induced cell injury in the study model.

A Potential Application of Triangular Microwells to Entrap Single Cancer Cells: A Canine Cutaneous Mast Cell Tumor Model

“…Fundamentally, the deleterious effects of a microfluidic system depend on the designed geometry [7], the intrinsic hydrodynamic mechanisms used, and the cell type. For example, the inertial hydrodynamic forces have been shown to be detrimental to sorted MCT cells, but not to canine leukocytes, in the case of size-based cell separation with a spiral microchannel [24].…”

Investigation of Leukocyte Viability and Damage in Spiral Microchannel and Contraction-Expansion Array

“…The specimen was mounted onto stubs with conductive tape and coat with gold (sputter coater, Balzers model SCD 040, Germany). Finally, they were observed by means of SEM [37]. In this study, we define “normal cell” to refer to cells in a circular shape.…”

Hydrodynamic dissection ofStentor coeruleusin a microfluidic cross junction

“…56 Hydrodynamic stresses are also known to damage cells and reduce their viability, but the extent of damage depends on the magnitude of shear stresses and the cell type. 43,[57][58][59][60] Hydrodynamic stresses ranging from ∼0.1 Pa to 100 Pa have been used in previously reported studies to achieve membrane permeabilization. 39,43,44 For adherent mammalian cells, mechanical forces are transmitted from the integrins to the nucleus via the cytoskeletal fibers to trigger signaling pathways and can lead to cell death.…”