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      Haemopoietic Origin of Myofibroblasts Formed in the Peritoneal Cavity in Response to a Foreign Body


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          This study utilized both in vivo and in vitro techniques to investigate whether cells of bone marrow origin can differentiate into smooth muscle-like cells (myofibroblasts) with contractile filaments and proteins. Female C57BL/6 mice expressing the Ly5.2 antigen on the surface of their haemopoietic cells had four pieces of silastic tubing (3 × 0.5 mm outer diameter) or boiled blood clot (2–3 mm diameter) placed in their peritoneal cavity. After 3, 5, 7 and 14 days (n = 4/group) the implants were removed and those that had remained free-floating were processed for light microscopy, immunohistochemistry and electron microscopy. In the first 3–5 days, rounded cells adhered to the entire surface of the tubing then flattened. These cells stained with fluoresceinated antibodies to Ly5.2 showing that they were derived from haemopoietic cells. By 14 days the cells had become elongated and multilayered in a collagen matrix, forming a thick tissue capsule around the tubing or boiled clot. They contained contractile filaments and stained with antibodies to α-smooth muscle actin but no longer stained for Ly5.2. A separate set of female C57BL/6 Ly5.2 mice were X-irradiated to destroy bone marrow then immediately transfused with 10<sup>6</sup> nucleated bone marrow cells taken from the femur and tibia of a congenic strain of male mice expressing the Ly5.1 allele. Eight of the female mice with successful engraftment (80–99%) had silastic tubing implanted in the peritoneal cavity. After 14 days, in situ hybridization with Y chromosome probe confirmed the male donor, and thus bone marrow, origin of the elongated cells that formed the capsule. In vitro studies showed that cells of the murine macrophage cell lines RAW 264.7 and J774 express α-smooth muscle actin after exposure to the cytokine γ-interferon in vitro. These data show that bone marrow-derived cells can differentiate into smooth muscle-like cells and raises the possibility that blood-derived cells may contribute to the development of fibro-proliferative vascular diseases such as atherosclerosis.

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          Subendothelial smooth muscle cells of human aorta express macrophage antigen in situ and in vitro.

          Cells bearing a smooth muscle cell marker--alpha-actin and a macrophage marker--CD68 antigen were immunocytochemically identified on 'en face' preparations of human aortic intima. Cells, expressing smooth muscle alpha-actin, macrophage CD68 antigen and both markers, i.e. smooth muscle cells possessing the macrophage antigen, were identified both in grossly normal aortic areas and in atherosclerotic lesions (fatty streaks and atherosclerotic plaques). CD68-positive smooth muscle cells were most common in the lipid-rich areas: fatty streaks and atherosclerotic plaque shoulders. Cells expressing smooth muscle alpha-actin and CD68 were also revealed in primary cultures prepared from grossly normal and atherosclerotic intima. Cells expressing both antigens were found in all examined cultures. The proportion of these cells in cultures from grossly normal areas and atherosclerotic plaques was similar: 14.5 +/- 4.1 and 14.6 +/- 4.8%, respectively. Cultures from fatty streaks had a higher content of cells expressing both antigens: 25.1 +/- 7.0%. Modified low density lipoprotein-induced intracellular lipid accumulation in cells cultured from grossly normal intima led to a three-fold increase in the number of cells sharing alpha-actin and CD68 antigen. Accumulation of latex beads by phagocytosis had a similar effect. It was suggested that in atherosclerotic lesions intracellular lipid accumulation and other stimulators of phagocytosis may provoke the expression of macrophage-associated antigen CD68 in settled cells of the subendothelial intima of human aorta.

            Author and article information

            J Vasc Res
            Journal of Vascular Research
            S. Karger AG
            October 2000
            02 October 2000
            : 37
            : 5
            : 364-371
            aCentre for Research in Vascular Biology, University of Queensland, Brisbane, Australia; bDivision of Cardiovascular Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
            25752 J Vasc Res 2000;37:364–371
            © 2000 S. Karger AG, Basel

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            Page count
            Figures: 6, References: 23, Pages: 8
            Research Paper


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