GSM5593338: AI.3.MF: AI treated Tumor Macrophage Cells Run 1 Rep3; Mus musculus; RNA-Seq
Tumor samples were collected immediately after surgery and submerged in tissue storage solution and were dissociated using the mouse tumor dissociation kit. Briefly, tumors were cut into small pieces (~ 2-4 mm3) and transferred to a gentle MACS C tube containing 2.35 ml RPMI 1640, 100 μl Enzyme D, 50 μl Enzyme R and 12.5 μl Enzyme A. The program 37C_m_TDK_1 was used on a gentle MACS Octo Dissociator with Heaters to fully dissociate tumor tissue. After digestion, the solution was diluted in 10 ml RPMI 1640 and filtered through a prewetted 70 μm filter. Cells were pelleted and used for positive selection of immune cells using mouse CD45 MicroBeads. RNA libraries were prepared for sequencing using SMART-Seq v2 protocol for Illumina