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Tracking single hematopoietic stem cells in vivo using high-throughput sequencing in conjunction with viral genetic barcoding.
- Source :
-
Nature biotechnology [Nat Biotechnol] 2011 Oct 02; Vol. 29 (10), pp. 928-33. Date of Electronic Publication: 2011 Oct 02. - Publication Year :
- 2011
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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.
- Subjects :
- Animals
Cell Differentiation genetics
Cell Differentiation radiation effects
Cell Lineage genetics
Cell Lineage radiation effects
Clone Cells
Gene Library
Hematopoietic Stem Cells radiation effects
Lentivirus radiation effects
Mice
Mice, Inbred C57BL
Radiation
Sequence Analysis, DNA
DNA Barcoding, Taxonomic methods
Hematopoietic Stem Cells cytology
Hematopoietic Stem Cells virology
High-Throughput Nucleotide Sequencing methods
Lentivirus genetics
Single-Cell Analysis methods
Subjects
Details
- Language :
- English
- ISSN :
- 1546-1696
- Volume :
- 29
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Nature biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 21964413
- Full Text :
- https://doi.org/10.1038/nbt.1977