ELISA was performed using human VEGF Quantikine PharmPak kit for VEGF‐A (R and D systems, USA), by following the manufacturer's instructions. 100 μL of CMs was added to 96‐well microwells and incubated for 2 hours at room temperature. After washing with PBS, 100 μL of biotin‐conjugate was added and washed. Next, 100 μL of streptavidin‐HRP was added to each well and kept for 1 hour at room temperature, and 100 μL later mixed and incubated for 30 minutes. The reaction was terminated by the addition of 100 μl of terminating solution. Colour density was read at 450 nm using a microwell reader (BioTek, Winooski, VT, USA).

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