Wound Healing

Application note: A novel quantitative wounding assay

Cellular wounding assays are used to understand the cellular mechanisms of healing,
including cell signaling, migration, and proliferation. Most wounding assays, such as scratch wound assays, are performed using a mechanical method, such as using a pipette tip to induce a break in a confluent monolayer of cells. Although mechanical wounding physically damages the monolayer, it creates highly variable wound sizes and shapes and causes spilling of intracellular contents, all of which complicate the interpretation of mechanistic data. Download this application note to learn a method to enzymatically wound cells, thereby eliminating the majority of off-target cell damage and creating wounds with consistent size and shape.

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Scratch wound assays are commonly performed to assess cellular migration and cell proliferation, often in response to therapeutic drug application. Wounding assays are also used to further understand the molecular phenomena related to wound healing, which includes cell signaling, immune and healing response to bacterial and fungal infections, and tissue remodeling. Most wounding assays are performed using a mechanical means to generate a break in a confluent monolayer of cells. Often this involves drawing a pipet tip or a pin over the monolayer. This physically damages the monolayer as desired, but also damages the cells, causing contents to leak and potentially complicating the interpretation of data. The other drawback of this method is that wound size and shape is highly variable contributing to difficultly in data analysis and inability to directly compare experimental conditions.

The BioFlux system can be used to wound cells enzymatically. The microfluidic channels in the BioFlux plate are identical within <5%, conferring the ability to generate wounds by fluid flow that are highly regular in shape and similar in size. Migration and proliferation following wounding can be analyzed by microscopy using promoters of cell migrations, epidermal growth factors, and motility and cell cycle inhibitors.

Key BioFlux Advantages For Wound Healing Studies

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figure2 results fc modified hit like monoclonal antibody
Featured Article

Fc-modified HIT-like monoclonal antibody as a novel treatment for sepsis

Sepsis leads to multiorgan dysfunction and is associated with increased morbidity and mortality, worldwide. During sepsis, neutrophils release extracellular traps (NETs) to capture pathogens, however, NETs have also been found to contribute to tissue damage.

In this featured article, Gollomp et al. examined potential therapeutics aimed at reducing sepsis-related tissue damage induced by NETs. To accomplish their goal, the authors developed and deployed recombinant immunoglobulin G2bk anti-hPF4/heparin monoclonal antibody (hPF4). Human umbilical vein endothelial cells (HUVECs) were cultured and activated in the BioFlux system, allowing the investigators to examine the impact of hPF4 treatment on NET entrapment.
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Wound healing assay in BioFlux 24-well plate

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This is a wound healing assay performed using the 24 well dual flow plates. After a monolayer of HUVEC cells is seeded, the growth media remains in inlet A (top) while trypsin is introduced in inlet B (bottom) removing half of the cells. Growth media is then introduced and the rate of cell proliferation is measured.