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

R Lu, NF Neff, SR Quake, IL Weissman - Nature biotechnology, 2011 - nature.com
Nature biotechnology, 2011nature.com
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 …
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.
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