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. 2010 Sep;48(9):563-7.
doi: 10.1002/dvg.20654.

The contribution of the Tie2+ lineage to primitive and definitive hematopoietic cells

Yuefeng Tang  1 , Anne Harrington, Xuehui Yang, Robert E Friesel, Lucy Liaw
Affiliations

The contribution of the Tie2+ lineage to primitive and definitive hematopoietic cells

Yuefeng Tang et al. Genesis. 2010 Sep.

Abstract

The regulatory elements of the Tie2/Tek promoter are commonly used in mouse models to direct transgene expression to endothelial cells. Tunica intima endothelial kinase 2 (Tie2) is also expressed in hematopoietic cells, although this has not been fully characterized. We determine the lineages of adult hematopoietic cells derived from Tie2-expressing populations using Tie2-Cre;Rosa26R-EYFP mice. In Tie2-Cre;Rosa26R-EYFP mice, analysis of bone marrow cells showed Cre-mediated recombination in 85% of the population. In adult bone marrow and spleen, we analyzed subclasses of early hematopoietic progenitors, T cells, monocytes, granulocytes, and B cells. We found that ∼ 84% of each lineage was EYFP(+), and nearly all cells that come from Tie2-expressing lineages are CD45(+), confirming widespread contribution to definitive hematopoietic cells. In addition, more than 82% of blood cells within the embryonic yolk sac were of Tie2(+) origin. Our findings of high levels of Tie2-Cre recombination in the hematopoietic lineage have implications for the use of the Tie2-Cre mouse as a lineage-restricted driver strain.

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Figures

Figure 1
Figure 1. The contribution of Tie2+ lineage cells to adult blood cells
A) Representative FACS plots of staining of bone marrow cells for the hematopoietic markers CD45, CD4, CD8, CD41, Gr1, Mac1, B220, and Tie2 (x-axis) and EYFP (y-axis) from Tie2-Cre;Rosa26R-EYFP mice at 2 months of age. B) Summary data of multiple experiments (n=8) analyzing bone marrow cells from Tie2-Cre;Rosa26R-EYFP mice. Data are presented as the percentage of each lineage that was EYFP+ ± S.D. of the mean percentage of each lineage. C) Representative FACS plots of staining of spleen cells for the hematopoietic markers CD45, CD4, CD8, and B220 (x-axis) and EYFP (y-axis). Hematopoietic lineages analyzed represent T cells (CD4 and CD8), B cells (B220), granulocytes (Gr1), megakaryocytes (CD41), and macrophages (Mac1). D) Summary data of multiple experiments (n = 8) analyzing spleens from Tie2-Cre;Rosa26R-EYFP mice. Data are presented as the percentage of each lineage that are EYFP+ ± S.D. of the mean percentage of each lineage.
Figure 2
Figure 2. A Tie2+ origin for embryonic blood cells from Tie2-Cre;Rosa26R-EYFP embryos
A) Tie2-Cre;Rosa26R-EYFP or control Rosa26R-EYFP embryos were collected at E7.5. Arrow shows EYFP+ mesodermal aggregations in visceral yolk sac in regions of forming blood islands. B) Representative FACS plots of E9.5 Tie2-Cre;Rosa26R-EYFP yolk sacs stained for the primitive hematopoietic progenitor marker CD41 or erythroid cell marker Ter119 (x-axis) and EYFP (y-axis) to assess the origin of these lineages within the E9.5 yolk sac. C) Summary data of multiple experiments (n = 10) analyzing the yolk sacs of Tie2-Cre;Rosa26R-EYFP mice. Data are presented as the percentage of Ter119+ and CD41+ that are also EYFP+ ± S.D. of the mean percentage of each lineage.
Figure 3
Figure 3. The Tie2-Cre transgene is not active in circulating embryonic blood cells
Peripheral blood cells were collected from Tie2-CreER;Rosa26R-EYFP mice at E12.5 and induced in vitro with 4OTH for 24 hours in suspension. Cells were then analyzed for EYFP expression within the Ter119+ population. No significant expression of EYFP was observed.
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