A Tubing-Free Microfluidic Wound Healing Assay Enabling the Quantification of Vascular Smooth Muscle Cell Migration

Abstract

This paper presents a tubing-free microfluidic wound healing assay to quantify the migration of vascular smooth muscle cells (VSMCs), where gravity was used to generate a laminar flow within microfluidic channels, enabling cell seeding, culture, and wound generation. As the first systemic study to quantify the migration of VSMCs within microfluidic environments, the effects of channel geometries, surface modifications and chemokines on cellular migration were investigated, revealing that 1) height of the micro channels had a significant impact on cell migration; 2) the surface coating of collagen induced more migration of VSMCs than fibronectin coated surfaces and 3) platelet derived growth factor resulted in maximal cell migration compared to tumor necrosis factor alpha and fetal bovine serum. Furthermore, migrations of five types of VSMCs (e.g., the human vascular smooth muscle cell line, two types of primary vascular smooth cells, and VSMCs isolated from two human samples) were quantified, finding that VSMCs from the cell line and human samples demonstrated comparable migration distances, which were significantly lower than the migration distances of two primary cell types. As a platform technology, this wound healing assay may function as a new model to study migration of VSMCs within microfluidic environments.

Raising fluid walls around living cells

“…Wound-healing assays generally involve comparing cells growing in different conditions in different dishes ( 15 – 17 ), and various microfluidic devices have been developed for such assays ( 18 , 19 ). As many chambers can be built in one dish, and as liquid walls are easily destroyed, we exploit these features to monitor two wounds healing in one dish, in which different parts of the dish have been coated in different ways.…”

A microfluidics-based wound-healing assay for studying the effects of shear stresses, wound widths, and chemicals on the wound-healing process

“…A migration distance of around 600 μm was observed after treating cells with epidermal growth factor-contained culture medium for 24 hr. Other similar microfluidics-based assays were reported to study wound-healing processes of various cell types including vascular smooth muscle cells 24 , endothelial cells 25 , and alveolar epithelial-like cells (A549) 26,27 . However, all these assays were limited to creating only one wound in one experiment, and the external stimuli to be applied were limited to chemicals.…”

Proteomic Profiling Exosomes from Vascular Smooth Muscle Cell

“…We examined a widely used human VSMC cell line, T/G-HAVSMC, as the cell source. [10] In our initial studies, we no ticed that T/G-HA-VSMCs underwent apoptosis in the serum free condition. Under such conditions, apoptotic blebs were produced with sizes in the same range as exosomes.…”