Antibiotic and antimycotic

Bone Marrow-derived Mast Cell (BMMC) was obtained following the protocol described by (30 (link)). Briefly, bone marrow cells were obtained from femurs and tibias of 6-8-week-old C57BL/6 female mice. Cells were maintained in RPMI-1640 supplemented with 10% FBS, 5 μM β-mercaptoethanol (Life Technologies, USA) and 2% antibiotic and antimycotic (Sigma, USA) (complete RPMI 1640 medium) plus 10 ng/mL of murine recombinant IL-3 (Peprotech, USA) and 10 ng/mL of murine recombinant stem cell factor (Peprotech, USA)). Non-adherent cells were transferred to fresh culture medium twice a week for 6−9 weeks. The purity of BMMC was ≥90% assessed by flow cytometry after staining of CD117 (clone: 2B8, BioLegend, USA; 0.25 μg/mL) and FcϵRI (clone: MAR-1, BioLegend, USA; 0.16 μg/mL) (Supplementary Figure 1A).
Peritoneum-derived mast cells (PMC), cells were obtained from peritoneal cavity of mice and cultured in complete RPMI-1640 medium plus IL-3 (30 ng/mL) and SCF (20 ng/mL). Non-adherent cells were transferred to fresh culture medium twice a week for 3−4 weeks. The purity of PMC was ≥90% assessed by flow cytometry (Supplementary Figure 1C).
All experiments followed institutional biosecurity and safety procedures. All animal experiments were approved by the Research Ethics Committee of the ENCB, IPN (ZOO-016-2019).

A20 murine lymphoma cell line was purchased from the American Type Culture Collection (ATCC, TIB-208TM). A20 cells were cultured at 5% CO₂, 37 °C in RPMI-1640 Medium with L-glutamine and high-glucose (GIBCO Life Technologies) supplemented with 10% Fetal Bovine Serum (FBS, GIBCO Life technologies) and 1% Antibiotic and Antimycotic (Sigma-Aldrich). For in vitro studies and tumor induction, the cells were used at 85% confluence and the viability of the cells was always higher than 90%, as assessed by the trypan blue exclusion test.

Bone marrow cells were obtained from femurs and tibias of 6–8-week-old C57BL/6 female and male mice. Cells were maintained in RPMI-1640 supplemented with 10% FBS, 5 μM β-mercaptoethanol (Life Technologies, USA) and 2% antibiotic and antimycotic (Sigma, USA) (complete RPMI 1640 medium), plus 10 ng/mL of murine recombinant IL-3 (Peprotech, USA) and 10 ng/mL of murine recombinant stem cell factor (SCF) (Peprotech, USA). Non-adherent cells were transferred to fresh culture medium twice a week for 6 − 9 weeks. The purity of bone marrow-derived mast cells (BMMC) was ≥ 90% as assessed by flow cytometry after staining of CD117 (clone: 2B8, BioLegend, USA; 0.25 μg/mL) and FcεRI (clone: MAR R-1, BioLegend, USA; 0.16 μg/mL).