Single-cell gene expression profiles of the small intestine in aged mice from normal-fed and overfed litters

We employed an established neonatal overfeeding model by reducing litter size, assigning litters to either 4 pups per dam (Overfed group) or 8 pups per dam (Normal-fed control group). To systematically characterize the transcriptional impact of early-life overfeeding on intestinal epithelial cell heterogeneity in aged mice, we conducted single-cell RNA sequencing (scRNA-seq) of small intestinal tissues from 28-month-old mice from normal-fed and overfed litters. The 28-month-old pooled cohort was processed using the 10 x Genomics platform. Briefly, aged tissues were digested with Collagenase I/II and Neutral Protease, filtered, and depleted of dead cells. Single-cell libraries were generated using the Chromium Single Cell 3’ Reagent Kit v4 and sequenced on the Illumina NovaSeq™ X Plus platform by LC-Bio Technology Co., Ltd.