In Vitro Wound Healing
Epistem's in vitro wound healing assays generate rapid mechanistic data to determine the effect of novel therapeutic agents on the processes implicated in wound healing including cell proliferation and migration, wound contraction and angiogenesis. Assays are performed using:
- Cell lines
- Primary human or murine keratinocytes
- Primary human or murine fibroblasts
Primary cells can be provided from a variety of donor types and ages, or other species, on request.
Cell Proliferation and Migration Assays
The rapid closure of open wounds by the proliferation and migration of epidermal cells is vital to reduce the risk of infection and further complications. These assays can be used to screen for novel therapeutic agents that promote wound closure.
Scratch Assay - Monolayers of confluent cells are mechanically scratched and the rate at which the cells migrate to close the scratch is measured. Simultaneous analysis of effects on proliferation and migration can be performed.
High Throughput Migration Assay - Monolayers of cells are seeded into 96-well plates, with stoppers in the centre of each well creating a void. Stoppers are removed and the rate of cell migration into the void is quantified. Effects on proliferation can be measured simultaneously and used to determine the relationship with cell migration.
3D Migration Assay - A collagen window is created around a spheroid or dense ball of fibroblasts and suspended in media. Cell migration out from this ball of cells is calculated as a percentage of the original area.
Wound Contraction Assay
Promoting fibroblast contraction reduces the size of an open wound, speeding up the healing process. Epistem's Wound Contraction Assay can be used to screen for the effect of novel therapeutic agents on wound contraction.
Fibroblasts are seeded into collagen gels and the novel therapeutic agent applied. Contraction typically occurs over the following 5 days. Gels are imaged and the effect of novel therapeutic agent on the rate of contraction determined.
Human endothelial cells are cultured in conditions that permit tubule formation. Quantification of vessel length and branching in preformed via labelling with anti-CD31 antibody. (See our Angiogenesis section for further information)