Abstract
The nature of hematopoietic stem cells under normal hematopoiesis remained largely unknown due to the limited assays available to monitor their behavior in situ. Here, we develop a new mouse model to transfer genes specifically into the primitive hematopoietic stem cell compartment through the utilization of a modified Rcas/TVA system. We succeeded in transferring a GFP reporter gene into adult hematopoietic stem cells in vivo, which are predominantly quiescent, by generating pseudotyped-lentivirus. Furthermore, we demonstrate the utility of this system to study neonatal hematopoiesis, a developmental stage that has been difficult to analyze to date. Using the system developed in this study, we observed continuous multi-lineage hematopoietic cell supply in peripheral blood from Krt7-positive hematopoietic stem cells during unperturbed homeostatic condition. This powerful experimental system could provide a new standard tool to analyze hematopoiesis under physiological condition without transplantation.
Highlights
Hematopoietic stem cells (HSCs) are one of the most well-studied adult stem cell populations and have provided many important concepts in the stem cell biology field[1,2,3,4,5]
We focused on Krt[7] as a potential marker for HSCs, and by using the system developed in this study, we confirm long-term multi-lineage hematopoiesis from a Krt7-expressing adult cell population in vivo, suggesting that Krt7-positive cells exist within the HSC population and serve as a source of mature blood cells in native, unstressed conditions
To investigate whether any cytokeratin family members displayed HSC-specific gene expression, we undertook RT-PCR in different hematopoietic cell fractions
Summary
Hematopoietic stem cells (HSCs) are one of the most well-studied adult stem cell populations and have provided many important concepts in the stem cell biology field[1,2,3,4,5]. The generation of mice expressing the tva gene under the control of a tissue- or cell type-specific promoter and their in vivo transduction by injection of Rcas virus into mice have been reported[16,17,18,19].
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