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Peritoneal repairing cells: a type of bone marrow derived progenitor cells involved in mesothelial regeneration.

Carmona R, Cano E, Grueso E, Ruiz-Villalba A, Bera TK, Gaztambide J, Segovia JC, Muñoz-Chápuli R - J. Cell. Mol. Med. (2010)

Bottom Line: This was observed in the injured area as well as in the surrounding not-injured peritoneal surfaces.We suggest that PRC constitute a type of monocyte-derived cells, closely related with the tissue-repairing cells known as 'fibrocytes' and specifically involved in peritoneal reparation.Thus, our results constitute a synthesis of the different scenarios hitherto proposed about peritoneal regeneration, particularly recruitment of circulating progenitor cells and adhesion of free-floating coelomic cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biology, Faculty of Science, University of Málaga, Málaga, Spain.

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Antigen immunolocalization in adherent cells obtained from peritoneal lavages 48 hrs after injury of the peritoneal wall. (A–C) show cells obtained from normal BALB/c mice whereas (D)–(F) show cells from mice reconstituted with GFP-expressing bone marrow. (A) Virtually all the CD45+ cells show a perinuclear dot-like pattern of cytokeratin immunoreactivity. Note a strongly CD45–/cytokeratin+ immunoreactive cell, probably a delaminated mesothelial cell (arrow). (B) The perinuclear dot-like pattern of cytokeratin immunoreactivity is also present in mesothelial cells migrating from omentum explants, which were used as positive controls (arrows). Other cells still show an extended cytokeratin cytoskeleton (arrowhead). (C) Mesothelin immunoreactivity colocalized with CD68. Note the cytoplasmic and perinuclear CD68 localization. (D) Triple localization of mesothelin, cytokeratin and GFP. This immunostaining revealed different phenotypes, including triple positive cells (arrowheads), mesothelin+/cytokeratin+, GFP– cells which probably are delaminated mesothelial cells (white arrows) and GFP+, mesothelin–/cytokeratin– cells (yellow arrow). (E) The fibroblast marker FSP1 was expressed in many GFP+ cells (white arrows), as well as in GFP– cells (yellow arrows). Other GFP+ cells were negative for FSP1 (arrowhead). (F) Negative control incubated with isotype primary antibodies and with the same secondary antibodies as used in the rest of the figures.
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fig03: Antigen immunolocalization in adherent cells obtained from peritoneal lavages 48 hrs after injury of the peritoneal wall. (A–C) show cells obtained from normal BALB/c mice whereas (D)–(F) show cells from mice reconstituted with GFP-expressing bone marrow. (A) Virtually all the CD45+ cells show a perinuclear dot-like pattern of cytokeratin immunoreactivity. Note a strongly CD45–/cytokeratin+ immunoreactive cell, probably a delaminated mesothelial cell (arrow). (B) The perinuclear dot-like pattern of cytokeratin immunoreactivity is also present in mesothelial cells migrating from omentum explants, which were used as positive controls (arrows). Other cells still show an extended cytokeratin cytoskeleton (arrowhead). (C) Mesothelin immunoreactivity colocalized with CD68. Note the cytoplasmic and perinuclear CD68 localization. (D) Triple localization of mesothelin, cytokeratin and GFP. This immunostaining revealed different phenotypes, including triple positive cells (arrowheads), mesothelin+/cytokeratin+, GFP– cells which probably are delaminated mesothelial cells (white arrows) and GFP+, mesothelin–/cytokeratin– cells (yellow arrow). (E) The fibroblast marker FSP1 was expressed in many GFP+ cells (white arrows), as well as in GFP– cells (yellow arrows). Other GFP+ cells were negative for FSP1 (arrowhead). (F) Negative control incubated with isotype primary antibodies and with the same secondary antibodies as used in the rest of the figures.

Mentions: Colocalization of mesothelial and leucocytic markers was frequent in primary culture of cells obtained from peritoneal lavages 48 hrs after injury, either from mice with GFP-expressing bone marrow or from normal mice (Fig. 3). The colocalization was always found in large, spindle-shaped cells. Cytokeratin showed a characteristic perinuclear dot-like staining pattern (Fig. 3A, colocalization with CD45). This pattern is frequent in primary cultures of mesothelial cells, in which the cytokeratin cytoskeleton seems to collapse, as shown in Figure 3(B), where some mesothelial cells show also the perinuclear dot like pattern whereas others show a still well-developed cytokeratin cytoskeleton (arrowhead in Fig. 3B). CD68 showed an intracytoplasmic pattern and colocalization with mesothelin (Fig. 3C). In peritoneal cells obtained from mice with GFP-expressing bone marrow, GFP colocalized with both mesothelin and cytokeratin in some cells (arrowheads in Fig. 3D) whereas others were GFP–/cytokeratin+/mesothelin+ (white arrows in Fig. 3D). GFP also colocalized with the fibroblastic marker FSP1 in part of the cells (Fig. 3E, E’). Negative control was incubated with isotype IgG and the same secondary antibodies than in the other slides (Fig. 3F).


Peritoneal repairing cells: a type of bone marrow derived progenitor cells involved in mesothelial regeneration.

Carmona R, Cano E, Grueso E, Ruiz-Villalba A, Bera TK, Gaztambide J, Segovia JC, Muñoz-Chápuli R - J. Cell. Mol. Med. (2010)

Antigen immunolocalization in adherent cells obtained from peritoneal lavages 48 hrs after injury of the peritoneal wall. (A–C) show cells obtained from normal BALB/c mice whereas (D)–(F) show cells from mice reconstituted with GFP-expressing bone marrow. (A) Virtually all the CD45+ cells show a perinuclear dot-like pattern of cytokeratin immunoreactivity. Note a strongly CD45–/cytokeratin+ immunoreactive cell, probably a delaminated mesothelial cell (arrow). (B) The perinuclear dot-like pattern of cytokeratin immunoreactivity is also present in mesothelial cells migrating from omentum explants, which were used as positive controls (arrows). Other cells still show an extended cytokeratin cytoskeleton (arrowhead). (C) Mesothelin immunoreactivity colocalized with CD68. Note the cytoplasmic and perinuclear CD68 localization. (D) Triple localization of mesothelin, cytokeratin and GFP. This immunostaining revealed different phenotypes, including triple positive cells (arrowheads), mesothelin+/cytokeratin+, GFP– cells which probably are delaminated mesothelial cells (white arrows) and GFP+, mesothelin–/cytokeratin– cells (yellow arrow). (E) The fibroblast marker FSP1 was expressed in many GFP+ cells (white arrows), as well as in GFP– cells (yellow arrows). Other GFP+ cells were negative for FSP1 (arrowhead). (F) Negative control incubated with isotype primary antibodies and with the same secondary antibodies as used in the rest of the figures.
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Related In: Results  -  Collection

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fig03: Antigen immunolocalization in adherent cells obtained from peritoneal lavages 48 hrs after injury of the peritoneal wall. (A–C) show cells obtained from normal BALB/c mice whereas (D)–(F) show cells from mice reconstituted with GFP-expressing bone marrow. (A) Virtually all the CD45+ cells show a perinuclear dot-like pattern of cytokeratin immunoreactivity. Note a strongly CD45–/cytokeratin+ immunoreactive cell, probably a delaminated mesothelial cell (arrow). (B) The perinuclear dot-like pattern of cytokeratin immunoreactivity is also present in mesothelial cells migrating from omentum explants, which were used as positive controls (arrows). Other cells still show an extended cytokeratin cytoskeleton (arrowhead). (C) Mesothelin immunoreactivity colocalized with CD68. Note the cytoplasmic and perinuclear CD68 localization. (D) Triple localization of mesothelin, cytokeratin and GFP. This immunostaining revealed different phenotypes, including triple positive cells (arrowheads), mesothelin+/cytokeratin+, GFP– cells which probably are delaminated mesothelial cells (white arrows) and GFP+, mesothelin–/cytokeratin– cells (yellow arrow). (E) The fibroblast marker FSP1 was expressed in many GFP+ cells (white arrows), as well as in GFP– cells (yellow arrows). Other GFP+ cells were negative for FSP1 (arrowhead). (F) Negative control incubated with isotype primary antibodies and with the same secondary antibodies as used in the rest of the figures.
Mentions: Colocalization of mesothelial and leucocytic markers was frequent in primary culture of cells obtained from peritoneal lavages 48 hrs after injury, either from mice with GFP-expressing bone marrow or from normal mice (Fig. 3). The colocalization was always found in large, spindle-shaped cells. Cytokeratin showed a characteristic perinuclear dot-like staining pattern (Fig. 3A, colocalization with CD45). This pattern is frequent in primary cultures of mesothelial cells, in which the cytokeratin cytoskeleton seems to collapse, as shown in Figure 3(B), where some mesothelial cells show also the perinuclear dot like pattern whereas others show a still well-developed cytokeratin cytoskeleton (arrowhead in Fig. 3B). CD68 showed an intracytoplasmic pattern and colocalization with mesothelin (Fig. 3C). In peritoneal cells obtained from mice with GFP-expressing bone marrow, GFP colocalized with both mesothelin and cytokeratin in some cells (arrowheads in Fig. 3D) whereas others were GFP–/cytokeratin+/mesothelin+ (white arrows in Fig. 3D). GFP also colocalized with the fibroblastic marker FSP1 in part of the cells (Fig. 3E, E’). Negative control was incubated with isotype IgG and the same secondary antibodies than in the other slides (Fig. 3F).

Bottom Line: This was observed in the injured area as well as in the surrounding not-injured peritoneal surfaces.We suggest that PRC constitute a type of monocyte-derived cells, closely related with the tissue-repairing cells known as 'fibrocytes' and specifically involved in peritoneal reparation.Thus, our results constitute a synthesis of the different scenarios hitherto proposed about peritoneal regeneration, particularly recruitment of circulating progenitor cells and adhesion of free-floating coelomic cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biology, Faculty of Science, University of Málaga, Málaga, Spain.

Show MeSH
Related in: MedlinePlus