Single-cell whole-genome sequencing reveals the functional landscape of somatic mutations in B lymphocytes across the human lifespan

In order to establish a causal relationship between somatic mutations and aging, mutational events must be directly identified in primary human tissues. Single cell sequencing holds promise for the detection of a full complement of mutations in somatic cells, overcoming the challenges that arise due to the random nature and very low abundance of most somatic mutations; however, the genome amplification procedures required for single cell genomics have a high error rate. To address this problem, Vijg, et al. developed a highly accurate single cell multiple displacement amplification (SCMDA) to comprehensively determine the full spectrum of base substitutions in a single somatic cell, and thereby assess mutation accumulation as a function of age in human B lymphocytes from healthy individuals. To aid in this analysis, bulk B lymphocytes were isolated from PBMCs and subsequently plated on gelatin coated CytoSort™ Arrays. Using the CellRaft┬« Technology, single B lymphocytes were isolated and collected in PCR tubes for SCMDA and downstream analysis of somatic mutations.