Tracking single hematopoietic stem cells in vivo using high-throughput sequencing in conjunction with viral genetic barcoding

Nat Biotechnol. 2011 Oct 2;29(10):928-33. doi: 10.1038/nbt.1977.

Abstract

Disentangling cellular heterogeneity is a challenge in many fields, particularly in the stem cell and cancer biology fields. Here we demonstrate how to combine viral genetic barcoding with high-throughput sequencing to track single cells in a heterogeneous population. We use this technique to track the in vivo differentiation of unitary hematopoietic stem cells (HSCs). The results are consistent with single-cell transplantation studies but require two orders of magnitude fewer mice. In addition to its high throughput, the high sensitivity of the technique allows for a direct examination of the clonality of sparse cell populations such as HSCs. We show how these capabilities offer a clonal perspective of the HSC differentiation process. In particular, our data suggest that HSCs do not equally contribute to blood cells after irradiation-mediated transplantation, and that two distinct HSC differentiation patterns co-exist in the same recipient mouse after irradiation. This technique can be applied to any virus-accessible cell type for both in vitro and in vivo processes.

Publication types

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

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cell Differentiation / radiation effects
  • Cell Lineage / genetics
  • Cell Lineage / radiation effects
  • Clone Cells
  • DNA Barcoding, Taxonomic / methods*
  • Gene Library
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / radiation effects
  • Hematopoietic Stem Cells / virology*
  • High-Throughput Nucleotide Sequencing / methods*
  • Lentivirus / genetics*
  • Lentivirus / radiation effects
  • Mice
  • Mice, Inbred C57BL
  • Radiation
  • Sequence Analysis, DNA
  • Single-Cell Analysis / methods*