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Culture medium associated changes in the core proteome of macrophages and in their responses to copper oxide
Bastien Dalzon1,2,3, Hélène Diemer4,5, Véronique Collin-Faure1,2,3
1Laboratory of Chemistry and Biology of Metals, BIG/CBM, CEA Grenoble, Grenoble, France.
Abstract:
The physiology of cells cultured in vitro depends obviously on the external conditions, including the nutrients present in the culture medium. In order to test the influence of this parameter, J774 macrophages grown either in RPMI or in DMEM were compared by a combination of targeted analyses and a proteomic approach. The two media differ in their glucose, amino acids, and vitamins concentrations, but there were no significant differences in the cell cycle or in the percentage of phagocytic cells in both media, although the phagocytic capacity (i.e. the number of phagocytized particles) was higher in DMEM. Conversely, we found that J774 cells grown in RPMI produced more nitric oxide in response to lipopolysaccharide. The proteomic study highlighted differences affecting the central metabolism and nucleotide metabolism, cytoskeleton, protein degradation, and cell signaling. Furthermore, proteomics showed that J774 cells grown in RPMI or in DMEM and exposed to copper oxide nanoparticles respond rather differently, with only a few proteins similarly modulated between cells grown in both media. Taken together, our results show that the basal state of cells grown in two different media is different, and this may affect the way they respond to an external stimulus or stress.
Insights
Cell culture media significantly impacts macrophage physiology. Differences in nutrient composition between RPMI and DMEM affect J774 macrophage metabolism, nitric oxide production, and nanoparticle response, influencing cellular behavior.
Area of Science:
- Cell Biology
- Immunology
- Proteomics
Background:
- In vitro cell physiology is influenced by culture medium composition.
- J774 macrophages are a relevant model for studying immune cell responses.
Purpose of the Study:
- To investigate the impact of two common cell culture media (RPMI and DMEM) on J774 macrophage physiology.
- To compare the basal state and response to stimuli of macrophages cultured in different media.
Main Methods:
- Comparative analysis of J774 macrophages cultured in RPMI versus DMEM.
- Utilized targeted analyses and a proteomic approach.
- Assessed cell cycle, phagocytosis, nitric oxide production, and protein expression.
Main Results:
- No significant differences in cell cycle or phagocytic cell percentage, but higher phagocytic capacity in DMEM.
- RPMI-cultured cells produced more nitric oxide upon lipopolysaccharide stimulation.
- Proteomics revealed differential modulation of metabolism, cytoskeleton, and signaling pathways.
- Differential responses to copper oxide nanoparticles were observed based on culture medium.
Conclusions:
- The basal physiological state of J774 macrophages differs significantly between RPMI and DMEM.
- Culture medium choice influences macrophage response to external stimuli and nanoparticles.
- These findings highlight the importance of considering culture medium composition in in vitro immunological studies.
