Easysep mouse cd4 and cd8 t cell enrichment kits

For the isolation of T cells, the spleens from the treated and tumor-bearing C57BL/6 mice were harvested on day 7, 14, 21, 28 and 90 after the treatment (n=3 mice per group at each time point), and prepared using GentleMACS dissociator (Miltenyi Biotec). The CD4⁺ and CD8⁺ T cells were purified from splenocytes using Easysep mouse CD4⁺ and CD8⁺ T cell Enrichment Kits (StemCell Technologies, Vancouver, BC, Canada) according to the manufacturer's instructions. The CD4⁺ and CD8⁺ T cells with a purity of >90% were used for experiments.

For isolation of T cells, spleens from the treated and tumor-bearing C57BL/6 mice were collected on day 5 and 14 after the treatment (n = 4 mice per group at each time point), respectively, and splenocytes were prepared using GentleMACS™ dissociator (Miltenyi Biotec) and passed through a 40 μm nylon filter. CD4⁺ and CD8⁺ T cells were purified from splenocytes using Easysep mouse CD4⁺ and CD8⁺ T cell Enrichment Kits (StemCell Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions. CD4⁺ and CD8⁺ T cells with a purity of 90% were used for experiments. DCs were isolated by DC isolation micro-bead kit (Easysep^TM CD11c positive selection kit, StemCell Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions. For CD68⁺ macrophage isolation, splenocytes were plated in Dulbecco’s modified Eagle’s medium supplemented with 10% FBS at 37 °C in a humidified 5% CO₂ incubator for 1 h, then the supernatant fraction was poured off and the adherent splenocyte fraction was processed for CD68⁺ macrophage isolation by using EasySep^TM PE positive selection kit (StemCell Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions.