Halal Research Center of Islamic Republic of Iran. Tel.: +98-21-88909033 Fax: +98-21-88890858 Email: jhhhm@halal.ac.ir IR.MCIG No. 85915 Human, Health and Halal Metrics 2717-1027 5 1 2024 06 01 Effect of caffeine-treated mesenchymal stem cells' conditioned medium on murine breast cancer cell lines (4T1) 41 52 197263 10.30502/jhhhm.2024.446060.1085 EN Zahra Zeinali Department of Microbiology, Faculty of Veterinary Medicine, Division of Immunology, Urmia University, Iran. Leila Mahmoudzadeh Department of Microbiology, Faculty of Veterinary Medicine, Division of Immunology, Urmia University, Iran. 0000-0002-3432-421X Journal Article 2024 02 27 <strong>Background and objective: </strong>Multiple studies on cancer showed that Mesenchymal Stem Cells (MSCs) inherently have different effects on various tumors. MSCs respond to multiple factors such as Caffeine, Estrogen, Morphine, curcumin, Metformin, and LPS due to the presence of different receptors. This study investigated the effects of caffeine, a popular drink, on the crosstalk between MSCs and 4T1 cells.<br /><strong>Material and methods</strong><strong>: </strong>Mesenchymal stem cells were isolated from mice's tibia and femur bones. These cells were cultured for 14 days and subsequently treated with o as the control group, 0.1, 0.5, and 1 mM doses of caffeine for 24 h. Then, cells were cultured without fetal bovine serum (FBS) for 24 h, and the conditioned medium(CM) was isolated. The 4T1 cells were cultured with CM for 24 h, MTT reduction, and Neutral red uptake assay. The cytotoxicity and the rate of apoptosis and necrosis were detected by Acridin orang and PI staining. The presence of 4T1 cancer cells was detected using the flow cytometry method.<br /><strong>Results and conclusion</strong><strong>: </strong>The MSC treated with caffeine could have. The more cytotoxic effect on 4T1 cells compared to non-treated MSC was seen in a dose-dependent manner, such that in the group that received a 0.5 mM concentration of caffeine, the most significant impact was seen on cell line 4.1. This data showed that the CM of MSC, treated with caffeine, had a cytotoxic effect on 4T1 cells. Caffeine could suppress the growth rate of 4T1 cells. The ideal caffeine concentration is 0.5 mM. https://jhhhm.halal.ac.ir/article_197263_ca63ef7a32b18ab9d46e0ebc346325c9.pdf