Scratch Wound Healing assay

Introduction / Principle

Epidermal regeneration is a complex dynamic process intended to restore the cellular and structural components of the skin through the implementation of mechanisms of proliferation and migration of keratinocytes.

The follow-up of the healing process after injury of the cell monolayer is carried out by a automated cell imaging system: the IncuCyte S3 (SARTORIUS). This device allows to perform kinetics of cell migration in classical culture conditions under controlled atmosphere (37°C; 5% CO2) by quantifying cell confluence.              

WoundMaker 96 puits (SARTORIUS)

Protocol

Format96 wells
Cellular model 

HaCaT (CLS 300493)

Reference molecule FBS
Wounding of the cell monolayer at confluence with a WoundMaker (SARTORIUS) after weaning the cells
Incubation48H at 37°C under 5% CO2 
possibility to adapt the incubation time
AnalysisIncucyte® S3 analysis (SARTORIUS)
→ % Relative Wound Density

Notes :

  • Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration)

Recommendation :

  • Ensure solubility of the sample in a solvent compatible with cell culture

Monitoring of wound healing on HaCaT cells

FBS 0% / 48h treatment
FBS 0.1% / 48h treatment
FBS 0.5% / 48h treatment
FBS 1% / 48h treatment