ICELL8 SMART-Seq single-cell RNA-seq of artificial macrophage-tumor doublets and fusion-derived hybrid controls

This study profiles tumor-macrophage hybrid cells in colorectal cancer using single-cell transcriptomic and epigenomic approaches. Fusion-derived hybrid cells were generated by co-culture of MC38-H2B-RFP colorectal cancer cells with beta-actin-GFP bone marrow-derived macrophages, and patient-derived hybrid cells were isolated from primary colorectal tumors and peripheral blood by fluorescence-activated cell sorting based on co-expression of epithelial (EpCAM and/or ECAD) and immune (CD45) markers. The study compares hybrid cells with parental tumor and immune controls and includes artificial macrophage-tumor doublets generated on the ICELL8 platform to distinguish bona fide hybrids from technical doublets. This submission contains an ICELL8 / SMART-Seq Pro single-cell RNA-seq library used to compare fusion-derived hybrid cells with imaging-validated artificial macrophage-tumor doublets and parental controls. Hybrid cells, parental MC38 tumor cells, and first-passage macrophages were isolated by flow cytometry. Artificial doublets were generated on the ICELL8 platform by first dispensing macrophages to 3/4 of the chip and hybrids to the remaining 1/4, then swapping source plates and dispensing MC38 cells only into the macrophage-loaded wells. Wells were imaged after each dispense and classified by DAPI, GFP, and RFP signal. All downstream first-strand synthesis, cDNA amplification, tagmentation, indexing, and purification steps followed the Takara SMART-Seq Pro workflow.