engleski

ENERGY STATUS IN CELL SPHERES DEPENDING ON CELL ORIGIN, SIZE AND NUTRIENT AVAILABILITY

Unique cell properties and cell growth conditions directly influence the energy status of the cell, while the energy status itself directly affects the metabolic activity that supports or supresses a specific cell function. Information about the energy status of the cell spheroids can provide a valuable input for further research of the metabolic activity of tissue-like models. The goal of our research was to determine how the change in the cell growth conditions affects the changes in the energy status of the cell. The evaluation was focused on the three main parameters, namely the sphere size, the nutrient availability from the growing medium and the cell type used to form the spheres. Two different cell lines were used to form the spheres: the normal MRC-5 cell line and the glioblastoma D54 cell line. Spheroids were grown under different controlled nutrient availability conditions and different time points in low adherent plates with V-bottom. Cells in the spheres were labelled with TMRE dye for detection of changes in mitochondrial membrane potential. We used the colorimetric ATP assay to determine the levels of ATP in cells lysate, and the colorimetric LDH assay to measure the enzyme activity of the lactate dehydrogenise. The obtained results indicate that the energy status of the cells in the spheres is in direct correlation with the level of the nutrient source, while the dissociated spheres, changes in mitochondrial potential, ATP levels and enzyme activity indicated a deficit in the energy levels required for the normal cell function.

energy status, cell spheres, ATP, mitochondrial potential

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