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Arrayed lentiviral barcoding for quantification analysis of hematopoietic dynamics.

Jeanne Grosselin1, Karine Sii-Felice, Emmanuel Payen

  • 1CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France; Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France.

Stem Cells (Dayton, Ohio)
|April 5, 2013
PubMed
Summary

This study introduces arrayed lentiviral barcoding to track individual cell fates in vivo, overcoming limitations of previous methods. The new technique clarifies hematopoietic stem cell behavior and reveals common myeloid-lymphoid biases in blood formation.

Keywords:
HematopoiesisHematopoietic stem cellsIn vivo trackingLentiviral vector

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Area of Science:

  • Cell Biology
  • Genetics
  • Biotechnology

Background:

  • High-throughput DNA sequencing enables individual cell marking using DNA barcodes.
  • Existing nonarrayed DNA barcode methods face statistical biases and cannot track cell fate after ex vivo manipulation and in vivo pooling.
  • Understanding system dynamics of mixed cell populations requires precise cell tracking.

Purpose of the Study:

  • To develop and validate an arrayed lentiviral library of DNA barcodes for precise in vivo cell tracking.
  • To overcome limitations of nonarrayed barcoding methods, including statistical biases and inability to deconvolute cell fate after ex vivo exposure.
  • To apply the arrayed barcoding method to study hematopoietic stem cell behavior and regeneration.

Main Methods:

  • Derivation of an arrayed lentiviral library of DNA barcodes.
  • Proof-of-concept study involving hematopoietic regeneration after engraftment of mice with genetically modified autologous cells.
  • Quantification of hematopoietic regeneration using the arrayed barcoding system.

Main Results:

  • Demonstrated the resolving capacity of arrayed lentiviral barcoding in a mouse model.
  • Clarified the dynamics of hematopoietic stem cell behavior, bridging clonal-succession and continuous-recruitment models.
  • Revealed prevalent myeloid-lymphoid biases in steady-state hematopoiesis.

Conclusions:

  • Arrayed lentiviral barcoding is a versatile and powerful tool for deconvoluting in vivo cell dynamics.
  • The method offers significant applications in hematology, embryology, and cancer biology.
  • This approach enhances the understanding of complex biological systems at the single-cell level.