Evaluating Phytochemical Effects on Chicken Embryo Cells

ATCC CRL-1590 chicken embryo cells were grown and maintained in Dulbecco’s modified Eagle’s medium (DMEM; VWR Life Science, NY, United States) containing 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Carlsbad, CA, United States) and 5% tryptose phosphate broth (Sigma-Aldrich). The effect of phytochemicals on the viability of chicken embryo cells was determined using alamarBlue assay as described previously (Wagle et al., 2020a (link)). Briefly, cells were cultured in 96-well tissue culture plates (Costar) at ∼10⁵ cells per well to form a monolayer at 37°C in a humidified, 5% CO₂ incubator for 24 h. The monolayer was washed three times and treated with DMSO in DMEM, or ethanol in DMEM, or phytochemicals at SIC in DMEM or DMEM (control) for 2 h. Negative controls without chicken embryo cells containing DMEM were also included. AlamarBlue reagent was added to each well, including blanks and negative controls, and incubated at 37°C for 1 h. The fluorescence was read before and after incubation using Cytation 5 multimode reader (BioTek Instruments, Inc., Winooski, VT, United States) at 560/590 nm (excitation/emission).