Single-cell RNA-sequencing of tumor-infiltrating cells from intra-tibial injection of LLC1 cells in murine model.

We injected LLC1 cells into tibia to construct a murine model to study bone microenvironment in cancer bone metastasis. We used single cell RNA sequencing (scRNA-seq) to analyze the cellular heterogeneity of tumor-infiltrating cells. LLC1 cells were injected intra-tibially into CAG-ZsGreen mice. 14 days later tumor-bearing tibia were dissected and enzymatically dissociated into single cell suspension. Tumor-infiltrating cells were isolated by Fluorescence-activated cell sorting (FACS) according to the presence of ZsGreen signal and analyzed using scRNAseq.