Single-cell RNA sequencing of multi-lineage differentiating stress-enduring (Muse) cells in the naïve state and after phagocytosis of cardiomyocyte-, neural cell-, or hepatic cell–derived fragments
Single-cell RNA sequencing of multi-lineage differentiating stress-enduring (Muse) cells in the naïve state and after phagocytosis of cardiomyocyte-, neural cell-, or hepatic cell–derived fragments
Human bone marrow-mesenchymal stem cells (h-BM-MSCs) were purchased from Lonza (Basel, Switzerland, PT-2501). SSEA-3(+) Muse cells were collected by fluorescence-activated cell sorting (FACS; BD FACSAria™ II cell sorter, Becton Dickinson, San Jose, CA, USA). mCherry- or GFP-h-Muse cells (1 × 105) were incubated for 3 days with m-cardiomyocyte DDCs, r-neural cell DDCs, and r-hepatic cell DDCs derived from 2 × 105 cells to make Phago-cardio-, Phago-neuro-, and Phago-hepa-Muse cells, respectively, then washed out and cultured in medium (DMEM + 10% FBS) for 4 days. Cells for all samples were stained with barcode tag-conjugated β2-microglobulin antibody and CD298 antibody (BioLegend), counted and multiplexed, and prepared for single-cell capture. Single-cell capture and cDNA synthesis were performed by the BD Rhapsody Single-Cell Analysis System according to the manufacturer's instructions.