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  • Optimizing Tumor-Stromal Exosome Dose for Enhanced Proliferation in MDA-MB-231 Breast Cancer Cells

  • Fateme hanifi,1 Mina ramezani,2,*
    1. Department of Biology, Central Tehran Branch, Islamic Azad University, Ashrafi Isfahani Highway, Imam Hassan Blvd., Tehran, Iran.
    2. Department of Biology, Central Tehran Branch, Islamic Azad University, Ashrafi Isfahani Highway, Imam Hassan Blvd., Tehran, Iran.


  • Introduction: Abstract Background and Aim: Exosomes shed by stromal cells within the tumor microenvironment are major players in intercellular communication and are known to influence the biological behavior of cancer cells, particularly their proliferation rates. This study set out to establish the optimal dose of tumor-derived stromal exosomes for further experimental use by assessing their effects on MDA-MB-231 breast cancer cell proliferation through the MTT assay.
  • Methods: MDA-MB-231 cells were grown to 80% confluency, trypsinized, and then seeded into 24-well plates at an optimized density. The cells received 24-hour treatments with exosome concentrations of 0 (control), 20, 30, and 40 µg/mL, each tested in duplicate. After adding the MTT reagent and dissolving the formazan crystals in DMSO, we read the absorbance at 490 nm using an ELISA reader. Proliferation rates were calculated as the fold change relative to the control group.
  • Results: Our findings showed that exosome concentrations of 20, 30, and 40 µg/mL boosted cell proliferation by 2.3-, 3.7-, and 2.5-fold, respectively, compared to the untreated controls. The strongest proliferative response was clearly seen at 30 µg/mL (3.7-fold increase; p < 0.01). Interestingly, when the dose was raised to 40 µg/mL, the proliferation rate dropped back to 2.5-fold, suggesting that higher exosome levels might introduce mild toxicity or inhibitory effects.
  • Conclusion: Based on these data, we selected 30 µg/mL as the ideal concentration of tumor-stromal exosomes for maximizing MDA-MB-231 cell proliferation, and it was consequently adopted for subsequent experimental steps. This dose-dependent biphasic pattern underscores the nuanced crosstalk between breast cancer cells and stromal-derived vesicles.
  • Keywords: Exosome, Extracellular vesicle,Tumor stromal cells,Breast cancer,MDA-MB-231 cell line,MTT assay,Cell

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