and cell proliferation
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.
Wound healing assay with trypsin
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. Cells are returned with normal growth media and the rate of growth is measured.
Key BioFlux Advantages For Wound Healing Studies
Achieve a wounding effect by using the two inlet one outlet well plate and trypsinizing half of the channel
Quantify wound size, cell migration, and cell proliferation
Generate tightly controlled wounds by enzymatic means and fluid flow
Cells at the leading edge of the wound remain intact; thus cytosolic leakage is not a factor in the interpretation of the data