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- Title
Tracking single hematopoietic stem cells in vivo using high-throughput sequencing in conjunction with viral genetic barcoding.
- Authors
Lu, Rong; Neff, Norma F; Quake, Stephen R; Weissman, Irving L
- 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
HEMATOPOIETIC stem cells; VIRAL genetics; CELL transplantation; CELL differentiation; IRRADIATION; LABORATORY mice
- Publication
Nature Biotechnology, 2011, Vol 29, Issue 10, p928
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt.1977