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Hematopoietic Origin of Murine Lung Fibroblasts.

McDonald LT, Mehrotra M, LaRue AC - Stem Cells Int (2015)

Bottom Line: Analysis of peripheral blood and lung tissues from engrafted mice demonstrated the ability of this population to give rise to CD45(+)/Discoidin-Domain Receptor-2(+) (DDR2) circulating fibroblast precursors (CFPs) in blood and fibroblast populations in lung.An HSC origin for lung fibroblasts was confirmed using a novel clonal cell transplantation method in which the bone marrow is reconstituted by a clonal population derived from a single HSC.Together, these findings provide evidence for an HSC contribution to lung fibroblasts and demonstrate a circulating intermediate through the CD45(+)/DDR2(+) HSC-derived CFP.

View Article: PubMed Central - PubMed

Affiliation: Research Services, Ralph H. Johnson Department of Veterans Affairs Medical Center, Charleston, SC 29401, USA ; Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29425, USA.

ABSTRACT
Multiple origins, including the bone marrow, have been suggested to contribute to fibroblast populations in the lung. Using bone marrow reconstitution strategies, the present study tested the hypothesis that the bone marrow hematopoietic stem cell (HSC) gives rise to lung tissue fibroblasts in vivo. Data demonstrate that the nonadherent bone marrow fraction is enriched for CD45(+) HSC-derived cells and was able to reconstitute hematopoiesis in lethally irradiated animals. Analysis of peripheral blood and lung tissues from engrafted mice demonstrated the ability of this population to give rise to CD45(+)/Discoidin-Domain Receptor-2(+) (DDR2) circulating fibroblast precursors (CFPs) in blood and fibroblast populations in lung. An HSC origin for lung fibroblasts was confirmed using a novel clonal cell transplantation method in which the bone marrow is reconstituted by a clonal population derived from a single HSC. Together, these findings provide evidence for an HSC contribution to lung fibroblasts and demonstrate a circulating intermediate through the CD45(+)/DDR2(+) HSC-derived CFP.

No MeSH data available.


Hematopoietic stem cells give rise to lung fibroblasts in vivo. Differential interference contrast (DIC), Hoechst nuclear stain (HO), EGFP, Collagen I, and DDR2 antibody stain are shown in Panels (a)–(e), respectively, from a representative section of lung tissue from a clonally engrafted mouse. Panel (f) shows merged image of GFP (green), Collagen I (red), and DDR2 (blue) staining. Panel (g) shows higher magnification of inset (boxed area) in Panel (f) with ∗ indicating a GFP/Collagen I/DDR2 expressing cell. Panels (h)–(n) depict images from secondary only staining controls.
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fig6: Hematopoietic stem cells give rise to lung fibroblasts in vivo. Differential interference contrast (DIC), Hoechst nuclear stain (HO), EGFP, Collagen I, and DDR2 antibody stain are shown in Panels (a)–(e), respectively, from a representative section of lung tissue from a clonally engrafted mouse. Panel (f) shows merged image of GFP (green), Collagen I (red), and DDR2 (blue) staining. Panel (g) shows higher magnification of inset (boxed area) in Panel (f) with ∗ indicating a GFP/Collagen I/DDR2 expressing cell. Panels (h)–(n) depict images from secondary only staining controls.

Mentions: As confirmation of the specific HSC origin of lung fibroblasts, mice engrafted with a clonal population derived from a single sorted HSC were generated as described previously [19–21, 25]. Total CD45.1−/EGFP+ engraftment and multilineage engraftment were confirmed (Table 3). Lungs were harvested and paraffin embedded and sections were stained for DDR2 and Collagen I to identify fibroblasts derived from the HSC. Numerous EGFP+ cells expressing DDR2 (28.8%) or Collagen I (13.5%) were identified (Figure 6 and quantified in Table 4), as were EGFP+ cells that expressed Collagen I and DDR2 (5.6%, representative cell indicated by ∗ in Figure 6) confirming an HSC origin of a population of lung fibroblasts.


Hematopoietic Origin of Murine Lung Fibroblasts.

McDonald LT, Mehrotra M, LaRue AC - Stem Cells Int (2015)

Hematopoietic stem cells give rise to lung fibroblasts in vivo. Differential interference contrast (DIC), Hoechst nuclear stain (HO), EGFP, Collagen I, and DDR2 antibody stain are shown in Panels (a)–(e), respectively, from a representative section of lung tissue from a clonally engrafted mouse. Panel (f) shows merged image of GFP (green), Collagen I (red), and DDR2 (blue) staining. Panel (g) shows higher magnification of inset (boxed area) in Panel (f) with ∗ indicating a GFP/Collagen I/DDR2 expressing cell. Panels (h)–(n) depict images from secondary only staining controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4491389&req=5

fig6: Hematopoietic stem cells give rise to lung fibroblasts in vivo. Differential interference contrast (DIC), Hoechst nuclear stain (HO), EGFP, Collagen I, and DDR2 antibody stain are shown in Panels (a)–(e), respectively, from a representative section of lung tissue from a clonally engrafted mouse. Panel (f) shows merged image of GFP (green), Collagen I (red), and DDR2 (blue) staining. Panel (g) shows higher magnification of inset (boxed area) in Panel (f) with ∗ indicating a GFP/Collagen I/DDR2 expressing cell. Panels (h)–(n) depict images from secondary only staining controls.
Mentions: As confirmation of the specific HSC origin of lung fibroblasts, mice engrafted with a clonal population derived from a single sorted HSC were generated as described previously [19–21, 25]. Total CD45.1−/EGFP+ engraftment and multilineage engraftment were confirmed (Table 3). Lungs were harvested and paraffin embedded and sections were stained for DDR2 and Collagen I to identify fibroblasts derived from the HSC. Numerous EGFP+ cells expressing DDR2 (28.8%) or Collagen I (13.5%) were identified (Figure 6 and quantified in Table 4), as were EGFP+ cells that expressed Collagen I and DDR2 (5.6%, representative cell indicated by ∗ in Figure 6) confirming an HSC origin of a population of lung fibroblasts.

Bottom Line: Analysis of peripheral blood and lung tissues from engrafted mice demonstrated the ability of this population to give rise to CD45(+)/Discoidin-Domain Receptor-2(+) (DDR2) circulating fibroblast precursors (CFPs) in blood and fibroblast populations in lung.An HSC origin for lung fibroblasts was confirmed using a novel clonal cell transplantation method in which the bone marrow is reconstituted by a clonal population derived from a single HSC.Together, these findings provide evidence for an HSC contribution to lung fibroblasts and demonstrate a circulating intermediate through the CD45(+)/DDR2(+) HSC-derived CFP.

View Article: PubMed Central - PubMed

Affiliation: Research Services, Ralph H. Johnson Department of Veterans Affairs Medical Center, Charleston, SC 29401, USA ; Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29425, USA.

ABSTRACT
Multiple origins, including the bone marrow, have been suggested to contribute to fibroblast populations in the lung. Using bone marrow reconstitution strategies, the present study tested the hypothesis that the bone marrow hematopoietic stem cell (HSC) gives rise to lung tissue fibroblasts in vivo. Data demonstrate that the nonadherent bone marrow fraction is enriched for CD45(+) HSC-derived cells and was able to reconstitute hematopoiesis in lethally irradiated animals. Analysis of peripheral blood and lung tissues from engrafted mice demonstrated the ability of this population to give rise to CD45(+)/Discoidin-Domain Receptor-2(+) (DDR2) circulating fibroblast precursors (CFPs) in blood and fibroblast populations in lung. An HSC origin for lung fibroblasts was confirmed using a novel clonal cell transplantation method in which the bone marrow is reconstituted by a clonal population derived from a single HSC. Together, these findings provide evidence for an HSC contribution to lung fibroblasts and demonstrate a circulating intermediate through the CD45(+)/DDR2(+) HSC-derived CFP.

No MeSH data available.