Single-Cell RNA Sequencing of Human Embryonic Stem Cell Differentiation Delineates Adverse Effects of Nicotine on Embryonic Development

Stem Cell Reports. 2019 Apr 9;12(4):772-786. doi: 10.1016/j.stemcr.2019.01.022. Epub 2019 Feb 28.


Nicotine, the main chemical constituent of tobacco, is highly detrimental to the developing fetus by increasing the risk of gestational complications and organ disorders. The effects of nicotine on human embryonic development and related mechanisms, however, remain poorly understood. Here, we performed single-cell RNA sequencing (scRNA-seq) of human embryonic stem cell (hESC)-derived embryoid body (EB) in the presence or absence of nicotine. Nicotine-induced lineage-specific responses and dysregulated cell-to-cell communication in EBs, shedding light on the adverse effects of nicotine on human embryonic development. In addition, nicotine reduced cell viability, increased reactive oxygen species (ROS), and altered cell cycling in EBs. Abnormal Ca2+ signaling was found in muscle cells upon nicotine exposure, as verified in hESC-derived cardiomyocytes. Consequently, our scRNA-seq data suggest direct adverse effects of nicotine on hESC differentiation at the single-cell level and offer a new method for evaluating drug and environmental toxicity on human embryonic development in utero.

Keywords: differentiation; embryonic development; embryonic stem cells; nicotine; single-cell RNA sequencing; smoking.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism
  • Cell Communication
  • Cell Cycle / genetics
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics*
  • Embryonic Development / genetics*
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / drug effects
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / metabolism
  • Nicotine / pharmacology*
  • Reactive Oxygen Species
  • Sequence Analysis, RNA
  • Single-Cell Analysis
  • Transcriptome


  • Reactive Oxygen Species
  • Nicotine
  • Calcium