ComProliM: A cell growth assay robust to initial cell number in co-culture system

Abstract

Cell growth is conventionally quantified using CCK-8 or MTT assays, but these methods display considerable sensitivity to initial cell quantities. Inherent sampling errors during cell counting and seeding make it impossible to achieve an absolute equivalence of initial cell numbers, potentially confounding the results of CCK-8 or MTT assays. In the present study, we introduce a novel cell proliferation assay, ComProliM, predicated on cell competition theory. Both numeral simulations and empirical testing demonstrate that ComProliM index (CPMI) reliably represents cell growth rate and is not influenced by variations in initial cell number. Intriguingly, two adherent cells of differing fluorescence states are co-cultured, suggesting that ComProliM can be successfully employed in co-culture system cell proliferation assays, including, for instance, the exploration of subclone interactions. We anticipate ComProliM will provide a viable alternative for quantifying adherent cell growth rates, particularly in cases where conventional methodologies deliver inconsistent or ambiguous results.