Functional division of insect blood cells in immunity by single-cell RNA sequencing

This study aimed to uncover the functional diversity and differentiation pathways of hemocytes in Spodoptera exigua by applying single-cell RNA sequencing (scRNA-seq). The objective was to classify hemocyte subtypes beyond traditional morphology and identify distinct gene expression profiles associated with immune responses, differentiation, and physiological functions. Fifth-instar Spodoptera exigua larvae were injected with Escherichia coli, incubated for 8 h, and hemolymph was collected for single-cell preparation. Cells were captured with 10x Chromium 3 prime v2, and libraries were prepared by poly(dT) priming with cell barcodes and UMIs, followed by Illumina paired-end sequencing. This sample represents hemocyte transcriptomes following immune activation.