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Related Concept Videos

Hematopoiesis01:21

Hematopoiesis

7.9K
The process of blood cell formation is called hematopoiesis. Hematopoiesis starts early during development, on the seventh day of embryogenesis. This phase of hematopoiesis is called the primitive wave, wherein the extraembryonic yolk sac allows the production of erythroid cells and endothelial cells from a common precursor called hemangioblast. The erythroid cells provide oxygen to support the growth of the rapidly dividing embryo. Hemangioblasts later develop into hematopoietic stem cells or...
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Multipotency of Hematopoietic Stem Cells01:19

Multipotency of Hematopoietic Stem Cells

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The hematopoietic stem cells or HSCs are multipotent, meaning they can differentiate and give rise to all blood and immune cells. HSCs are maintained in the quiescent stage until an external stimulus initiates their differentiation. The multipotent HSCs exist as two heterogeneous populations, long-term repopulating cells (LTRC) and short-term repopulating cells (STRC). The two HSC populations have different surface markers or receptors and are classified based on quiescence and long-term...
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Related Experiment Video

Updated: Dec 1, 2025

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
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Single-Cell Transcriptomic Analysis of Hematopoietic Cells.

Paulina M Strzelecka1,2,3,4, Anna M Ranzoni1,2,3, Ana Cvejic5,6,7

  • 1Wellcome Trust-Medical Research Council, Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, Cambridge, UK.

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2020
PubMed
Summary

Single-cell RNA sequencing (scRNA-Seq) offers unbiased analysis of cell transcriptomes, advancing hematology research. This study details protocols for Smart-Seq2 (plate-based) and 10× Genomics (droplet-based) methods for hematopoietic cells.

Keywords:
10× GenomicsDroplet-based RNA-SeqHematopoiesisOmicsPlate-based RNA-SeqSingle-cell RNA sequencingSingle-cell biologySingle-cell omicsSmart-Seq2Transcriptomics

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Area of Science:

  • Molecular Biology
  • Genomics
  • Hematology

Background:

  • Single-cell RNA sequencing (scRNA-Seq) provides comprehensive analysis of individual cell transcriptional states.
  • scRNA-Seq has significantly advanced the understanding of normal and malignant hematopoiesis over the last five years.
  • Various scRNA-Seq methods exist, all involving RNA to cDNA conversion and amplification for sequencing.

Purpose of the Study:

  • To describe detailed protocols for two prominent scRNA-Seq methods used in hematology research.
  • To compare plate-based and droplet-based scRNA-Seq approaches for hematopoietic cells.
  • To provide practical guidance for researchers designing scRNA-Seq experiments in hematopoiesis.

Main Methods:

  • Detailed protocols for Smart-Seq2 (plate-based) and 10× Genomics (droplet-based) scRNA-Seq are presented.
  • Both methods involve RNA extraction, cDNA synthesis, amplification, and subsequent sequencing.
  • The focus is on application to hematopoietic cells.

Main Results:

  • The study provides step-by-step protocols for implementing Smart-Seq2 and 10× Genomics platforms.
  • Advantages and disadvantages of droplet-based versus plate-based approaches are discussed in the context of experimental design.
  • The protocols are optimized for the analysis of hematopoietic cells.

Conclusions:

  • Smart-Seq2 and 10× Genomics represent widely used, yet distinct, scRNA-Seq methodologies for hematology.
  • Understanding the nuances of each platform (plate-based vs. droplet-based) is crucial for successful experimental design and data interpretation.
  • These detailed protocols facilitate the application of scRNA-Seq to further unravel hematopoiesis.