Dynamic Activity Analysis of Eukaryotic Cell Lines in the Evaluation of Plant Extracts

Plants are an inexhaustible source of new biologically active compounds that can be used to fight against various human diseases, including cancer. This study evaluates the effects of Pontederia crassipes leaf/petiole extracts on HDF and HEK293 cell lines using an integrated approach combining live-cell imaging (Celena X cell imager) and resazurin-based viability assays. Methanolic extracts were prepared via two methods: (A) prolonged extraction and (B) ultrasonication. Dynamic changes in cell count and surface area were quantified through time-lapse imaging over 24 h, with trendline slope analysis (regression coefficients) providing growth kinetics. Ethanolic extracts (Method A) were later included for comparative analysis. Additionally, methanolic extract was fractionated by HPLC, and GC-MS revealed a fairly wide range of phytochemical components in methanol extract fractions. Methanolic extracts (Method A) showed dose-dependent effects, with HEK293 cells more sensitive (proliferation inhibition at 12.5 μg/mL) than HDF (200 μg/mL). Ethanolic extracts inhibited HDF at 25–200 μg/mL and HEK293 at 12.5 μg/mL. The developed method using live cell imaging showed that the decrease in resazurin signals correlated with cytostasis (stable cell count) rather than cytotoxicity. The results highlight the importance of using this method in conjunction with endpoint assays to obtain more detailed information when analyzing bioactive compounds.