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Hematopoietic Origin of Murine Lung Fibroblasts.

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

Bottom Line: 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.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.


Transplantation of nonadherent bone marrow results in multilineage hematopoietic engraftment. Shown is a representative analysis of total (a) and multilineage (b) engraftment approximately 2 months after transplantation of nonadherent bone marrow cells. Isotype (top panels) forward scatter and side scatter gate is shown (left) with IgG-PE versus EGFP gate (right). Representative total engraftment is shown in Panel (a) (bottom panels) with forward scatter and side scatter gate (left) and PE and EGFP gates shown (right). (b) Representative multilineage engraftment analysis is depicted in Panel (b). The isotype IgG PE gate and the EGFP population gated on the IgG PE population are depicted in (i). Analysis of B cell (ii), T cell (iii), and granulocyte/macrophage (iv) populations demonstrates EGFP+ cells representing 82.5%, 36.5%, and 85.9% of each lineage, respectively. In all dot plots (i–iv), total populations are shown on the left, and EGFP+ cells within the gated population are shown on the right. See Table 1 for engraftment for all mice.
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fig2: Transplantation of nonadherent bone marrow results in multilineage hematopoietic engraftment. Shown is a representative analysis of total (a) and multilineage (b) engraftment approximately 2 months after transplantation of nonadherent bone marrow cells. Isotype (top panels) forward scatter and side scatter gate is shown (left) with IgG-PE versus EGFP gate (right). Representative total engraftment is shown in Panel (a) (bottom panels) with forward scatter and side scatter gate (left) and PE and EGFP gates shown (right). (b) Representative multilineage engraftment analysis is depicted in Panel (b). The isotype IgG PE gate and the EGFP population gated on the IgG PE population are depicted in (i). Analysis of B cell (ii), T cell (iii), and granulocyte/macrophage (iv) populations demonstrates EGFP+ cells representing 82.5%, 36.5%, and 85.9% of each lineage, respectively. In all dot plots (i–iv), total populations are shown on the left, and EGFP+ cells within the gated population are shown on the right. See Table 1 for engraftment for all mice.

Mentions: Mice were transplanted with bone marrow after 96 hours of selection for the nonadherent [32, 33], CD45 enriched fraction. Recipient CD45.1 mice were analyzed for total engraftment by flow cytometry 10 weeks after transplantation. High level engraftment (>50%) of CD45.1−/EGFP+ nucleated peripheral blood cells was confirmed. Total engraftment from a representative animal is depicted (Figure 2(a)). One characteristic of hematopoietic stem cells is in their capacity to repopulate all blood lineages. Therefore, as additional confirmation of enrichment of the nonadherent bone marrow fraction for HSCs [27, 29, 32–34], multilineage engraftment was confirmed in transplanted mice by flow cytometric analysis. Analysis revealed high level engraftment of EGFP+ cells in each of the blood lineages including the B cell (B220+), T cell (Thy1.2+), and granulocyte/macrophage (Gr1+/Mac1+) populations of nucleated peripheral blood cells at 10 weeks after transplant. Multilineage engraftment from a representative animal is shown (Figure 2(b)). Total and multilineage engraftment of each of the animals used herein is included in Table 1.


Hematopoietic Origin of Murine Lung Fibroblasts.

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

Transplantation of nonadherent bone marrow results in multilineage hematopoietic engraftment. Shown is a representative analysis of total (a) and multilineage (b) engraftment approximately 2 months after transplantation of nonadherent bone marrow cells. Isotype (top panels) forward scatter and side scatter gate is shown (left) with IgG-PE versus EGFP gate (right). Representative total engraftment is shown in Panel (a) (bottom panels) with forward scatter and side scatter gate (left) and PE and EGFP gates shown (right). (b) Representative multilineage engraftment analysis is depicted in Panel (b). The isotype IgG PE gate and the EGFP population gated on the IgG PE population are depicted in (i). Analysis of B cell (ii), T cell (iii), and granulocyte/macrophage (iv) populations demonstrates EGFP+ cells representing 82.5%, 36.5%, and 85.9% of each lineage, respectively. In all dot plots (i–iv), total populations are shown on the left, and EGFP+ cells within the gated population are shown on the right. See Table 1 for engraftment for all mice.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Transplantation of nonadherent bone marrow results in multilineage hematopoietic engraftment. Shown is a representative analysis of total (a) and multilineage (b) engraftment approximately 2 months after transplantation of nonadherent bone marrow cells. Isotype (top panels) forward scatter and side scatter gate is shown (left) with IgG-PE versus EGFP gate (right). Representative total engraftment is shown in Panel (a) (bottom panels) with forward scatter and side scatter gate (left) and PE and EGFP gates shown (right). (b) Representative multilineage engraftment analysis is depicted in Panel (b). The isotype IgG PE gate and the EGFP population gated on the IgG PE population are depicted in (i). Analysis of B cell (ii), T cell (iii), and granulocyte/macrophage (iv) populations demonstrates EGFP+ cells representing 82.5%, 36.5%, and 85.9% of each lineage, respectively. In all dot plots (i–iv), total populations are shown on the left, and EGFP+ cells within the gated population are shown on the right. See Table 1 for engraftment for all mice.
Mentions: Mice were transplanted with bone marrow after 96 hours of selection for the nonadherent [32, 33], CD45 enriched fraction. Recipient CD45.1 mice were analyzed for total engraftment by flow cytometry 10 weeks after transplantation. High level engraftment (>50%) of CD45.1−/EGFP+ nucleated peripheral blood cells was confirmed. Total engraftment from a representative animal is depicted (Figure 2(a)). One characteristic of hematopoietic stem cells is in their capacity to repopulate all blood lineages. Therefore, as additional confirmation of enrichment of the nonadherent bone marrow fraction for HSCs [27, 29, 32–34], multilineage engraftment was confirmed in transplanted mice by flow cytometric analysis. Analysis revealed high level engraftment of EGFP+ cells in each of the blood lineages including the B cell (B220+), T cell (Thy1.2+), and granulocyte/macrophage (Gr1+/Mac1+) populations of nucleated peripheral blood cells at 10 weeks after transplant. Multilineage engraftment from a representative animal is shown (Figure 2(b)). Total and multilineage engraftment of each of the animals used herein is included in Table 1.

Bottom Line: 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.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.