Cell viability was assessed by evaluating the membrane integrity using the Evans blue method. Therefore, viable cells (unstained) and dead cells (stained) were identified and counted on a Neubauer improved hemocytometer (Sigma-Aldrich) using an optic microscope (Zeiss Axio Imager Z1). The cell viability was defined as the percentage of viable cells relative to total cells (i.e., viable and dead cells). Cell growth rate was assessed every week using the cells counts with Evans blue method, as previously described in Ventura et al. (2018) (link) and Fricano et al. (2020) (link). The following formula was then used to calculate the weekly growth rate:
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