Single-cell sequencing in stem cell biology

Abstract

Cell-to-cell variation and heterogeneity are fundamental and intrinsic characteristics of stem cell populations, but these differences are masked when bulk cells are used for omic analysis. Single-cell sequencing technologies serve as powerful tools to dissect cellular heterogeneity comprehensively and to identify distinct phenotypic cell types, even within a 'homogeneous' stem cell population. These technologies, including single-cell genome, epigenome, and transcriptome sequencing technologies, have been developing rapidly in recent years. The application of these methods to different types of stem cells, including pluripotent stem cells and tissue-specific stem cells, has led to exciting new findings in the stem cell field. In this review, we discuss the recent progress as well as future perspectives in the methodologies and applications of single-cell omic sequencing technologies.

Fig. 1

Flowchart of a typical single cell omic sequencing project. A typical single-cell sequencing project comprises four major steps: experimental design, performing the experiments, bioinformatic analyses and verification of the discoveries. Here we use a single-cell RNA-seq project as an example. Note that if the project fails at any step, researchers should go back to previous steps to identify the cause of the failure and re-design accordingly. In a real project, this process may need to be repeated several times. FACS fluorescence-activated cell sorting, MACS magnetic-activated cell sorting, qPCR quantitative polymerase chain reaction