Wound Healing Assays in Well Plate–Coupled Microfluidic Devices with Controlled Parallel Flow

The migratory or proliferative responses mounted by wounded cell monolayers are important to drug discovery and drug safety testing, as well as to basic research across a number of disciplines, including stem-cell biology, cell biology, ophthalmology, endocrinology, microbiology, oncology, and developmental biology. Scratch wounding by mechanical means is the golden standard to achieve an appropriate model system in which to study these cellular reactions. The scratch wounding technique is plagued by wound size irregularity, release of cytosolic contents along the wound edge, and difficulty in scaling up to higher throughput screening. To address these issues, we developed a microfluidic device coupled to a well plate in which wounds were produced enzymatically using highly controlled laminar flow streams. Within the device, epithelial cells were cultivated and exposed to different compounds. Proliferation and migration were characterized by bright-field microscopy. Resulting wound size was highly uniform compared with reported variability of manual scratch wounding. The protocol for wounding was fully automated using customized software, and response to wounding was followed in real time by microscopy.