Wound Healing

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 mechanical means to generate a rift in a confluent monolayer of cells. Often this involves drawing a pipet tip or pin over the monolayer. This physically damages the monolayer as desired, but also damages the surrounding 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 employed to wound cells enzymatically with trypsin, which eliminates cell damage at the wound edge. This novel approach provides several benefits to running wound healing assays using BioFlux:

• Consistent and repeatable wound size created by fluid flow
• Direct imaging of the wounds using time-lapse microscopy
• Parallel format allows up to 96 simultaneous wound assays
• High content analysis including wound healing kinetics, cellular morphology and sub-cellular imaging

	Wounded rat lung epithelium (RLE) monolayer healing in serum rich medium in time-lapse over 6 hours.
	Wound response under migration (top) and proliferation conditions (bottom). RLE monolayers were wounded with trypsin and treated with medium containing serum (proliferation) or without serum (migration). Additional treatments such as EGF, which promotes migration, and cytochalsin D, which impedes migration and proliferation were tested simultaneously.