Hen’s Egg Test – Chorio-Allantoic Membrane

Method Description

This method is intented to enter a battery of assays to replace rabbits on Eye Irritation Test, as indication of non-irritant substances and a Bottom-Up strategy. Leghorn SPF eggs are used. The test substance is applied to the surface of the CAM. The CAM is evaluated by the appearance of following: congestion (hyperemia), hemorrhage (bleeding), and coagulation (intra- and extravascular). In Brazil, HET-CAM is already accepted by ANVISA as a screening test, but nor for regulatory purposes.