intra-single cell sequencing (iSCseq) spotlights transcriptomic and epigenetic heterogeneity inside multinucleated osteoclast

Multinucleated giant cells like osteoclasts (OCs) often contain >10 nuclei. This raises the question of whether nuclei residing in the same cytoplasm are all regulated similarly or whether they are heterogeneous, expressing a different set of genes regulated by nucleus-specific epigenetic mechanisms. We combined imaging of living cells with confocal microscope, picking of intracellular components inside a single cell using the Single Cellome™ System SS2000 (Yokogawa, Japan), allowing next generation sequencing at high resolution for mRNA (iscRNA-seq) and for DNA methylation (iscWGBS; whole genome bisulfite sequencing) of any cellular components including a single OC nucleus. Our results reveal that individual nuclei within the same cell are heterogeneous in terms of gene expression and epigenetic regulation, possibly affecting regional cell function.