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Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
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Mapping Human Pluripotent Stem Cell-derived Erythroid Differentiation by Single-cell Transcriptome Analysis.

Zijuan Xin1, Wei Zhang1, Shangjin Gong1

  • 1CAS Key Laboratory of Genome Science and Information, Beijing Institute of Genomics, Chinese Academy of Sciences/China National Center of Bioinformation, Beijing 100101, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.

Genomics, Proteomics & Bioinformatics
|July 20, 2021
PubMed
Summary
This summary is machine-generated.

Induced pluripotent stem cells (iPSCs) offer a solution for red blood cell (RBC) regeneration. This study maps the iPSC-derived erythroid differentiation pathway, identifying key transcription factors and cell stages for optimized RBC production.

Keywords:
Differentiation trajectoryErythropoiesisHematopoiesisiPSCscRNA-seq

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Area of Science:

  • Stem Cell Biology
  • Hematopoiesis
  • Genomics

Background:

  • Clinical demand for functional red blood cells (RBCs) exceeds supply, necessitating in vitro regeneration methods.
  • Induced pluripotent stem cells (iPSCs) present a viable alternative to cord blood and embryonic stem cells, avoiding ethical concerns and immune rejection.
  • A complete single-cell level differentiation pathway for iPSC-derived erythroid systems is currently lacking.

Purpose of the Study:

  • To establish and characterize an RBC regeneration system using the iPSC line BC1.
  • To map the cell lineage and differentiation trajectory of iPSCs during erythroid differentiation at single-cell resolution.
  • To identify regulatory transcription factor (TF) networks and cell-cell interactions governing this process.

Main Methods:

  • Utilized the 10X Genomics single-cell transcriptome platform to analyze iPSC differentiation on day 14.
  • Embryoid body (EB) culture was employed for iPSC differentiation.
  • Bioinformatic analysis was used to identify cell classifications, TF networks, and cell-cell contacts.

Main Results:

  • Observed unsynchronized iPSC differentiation, with cells resembling yolk sac hematopoiesis, including mesodermal and blood cells.
  • Identified six distinct cell classifications and characterized regulatory TF networks and cell-cell interactions.
  • Mapped erythroid cells at various stages (e.g., BFU-E, ortho-E) and found stage-specific TF regulation (e.g., TFDP1, FOXO3). Immune erythroid cells were also identified.

Conclusions:

  • Provides systematic theoretical guidance for optimizing iPSC-derived erythroid differentiation systems.
  • The established system serves as a valuable model for simulating in vivo hematopoietic development.
  • Highlights dynamic expression of cell adhesion molecules and estrogen-responsive genes during iPSC differentiation.