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Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Published on: January 7, 2020

A Modified SMART-Seq Method for Single-Cell Transcriptomic Analysis of Embryoid Body Differentiation.

Jianqun Zheng¹, Ying Ye², Qiushi Xu¹

¹Shenzhen Key Laboratory of Gene Regulation and Systems Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China.

Methods in Molecular Biology (Clifton, N.J.)

|October 18, 2021

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We developed a simple single-cell gene expression profiling method for mouse embryoid bodies (EBs) derived from mouse embryonic stem cells (mESCs). This technique reveals cellular identities during early embryo development.

Area of Science:

Developmental Biology
Stem Cell Biology
Genomics

Background:

Embryoid bodies (EBs) model early embryonic development by aggregating cells from pluripotent embryonic stem cells (ESCs).
Investigating the diverse cell types within EBs at a single-cell level is crucial for understanding developmental processes.

Purpose of the Study:

To present a robust and accessible method for single-cell gene expression profiling in differentiating mouse embryoid bodies (mEBs).
To enable detailed analysis of cellular heterogeneity and identity during EB formation.

Main Methods:

Adaptation of a SMART-seq-based protocol for accurate single-cell 3' transcript counting.
Utilizing standard molecular biology techniques and equipment for mouse ESCs (mESCs) differentiation into EBs.

Keywords:

Embryoid body Genomics Mouse embryonic stem cells Single-cell RNA-seq

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Integration with perturbation experiments for mechanistic investigations.

Main Results:

The described protocol provides high-resolution gene expression data at the single-cell level.
Accurate 3' counting facilitates the identification of distinct cell populations and their developmental trajectories within mEBs.
The method is shown to be effective for studying cellular identities during EB differentiation.

Conclusions:

This straightforward single-cell RNA sequencing method enhances the study of early embryonic development using EBs.
It offers a valuable tool for mechanistic studies of cell fate determination and differentiation in vitro.
The protocol supports detailed investigations into the cellular dynamics of embryogenesis.