Peripheral blood mononuclear cells pbmcs

Ovarian cancer cell lines were a kind gift from Dr. Ruby Huang (Cancer Science Institute, Singapore). HEK293, HFF1, and IMR90 were purchased from the ATCC. IGROV-1 was cultured in RPMI-1640 with 10% FBS. SKOV3, was cultured in a 1:1 ratio of DMEM (high glucose) and DMEM (low glucose) with 10% FBS added. IMR90, HEK293, and HFF-1 were cultured in DMEM (high glucose) supplemented with 10% FBS and 2 mM L-Glutamine.
Peripheral blood mononuclear cells (PBMCs) were purchased from StemCell Technologies. T cells were isolated from PBMCs using the EasySep™ Human T Cell Isolation Kit (StemCell Technologies, Vancouver, BC, Canada), according to the manufacturer’s protocol. In standard culture, T cells were activated with anti-CD3/CD28 Dynabeads (Life Technologies, Grand Island, NY, USA) at a 1:1 bead-to-cell ratio, and cultured in RPMI-1640 supplemented with 10% FBS, and a cytokine cocktail of IL-7 (20 U/mL), IL-15 (10 U/mL), and IL-21 (0.04 U/mL).

A panel of breast cancer cell lines with different expression of triple markers (ER, PR, HER2) was used for assay validation: MCF‐7, BT474, HCC1954, and MDA‐MB‐231. All cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Human breast cancer cell lines MCF‐7 and MDA‐MB‐231 were maintained in Dulbecco's modified Eagle's medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA); BT474 and HCC1954 were maintained in RPMI 1640 (Thermo Fisher Scientific, Waltham, MA, USA). All media were supplemented with 10% fetal bovine serum (FBS, Bio‐Techne Sales) and 1% penicillin‐streptomycin (Thermo Fisher Scientific, Waltham, MA, USA) and cultured at 37 °C with 5% CO₂. All cell lines were routinely tested using MycoAlert mycoplasma detection kit (Lonza, Basel, Switzerland). Peripheral blood mononuclear cells (PBMCs) were obtained from Stemcell Technologies (Cambridge, MA, USA).