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dc.contributor.authorGoan, Silvia-Renate
dc.contributor.authorJunghahn, Ilse
dc.contributor.authorWissler, Manuela
dc.contributor.authorBecker, Michael
dc.contributor.authorAumann, Jutta
dc.contributor.authorJust, Ursula
dc.contributor.authorMartiny-Baron, Georg
dc.contributor.authorFichtner, Iduna
dc.contributor.authorHenschler, Reinhard
dc.date.accessioned2010-01-19T16:45:40Z
dc.date.available2010-01-19T16:45:40Z
dc.date.issued2000-12-01
dc.identifier.citationDonor stromal cells from human blood engraft in NOD/SCID mice. 2000, 96 (12):3971-8 Blooden
dc.identifier.issn0006-4971
dc.identifier.pmid11090086
dc.identifier.urihttp://hdl.handle.net/10541/90055
dc.description.abstractLittle is known about the presence, frequency, and in vivo proliferative potential of stromal cells within blood-derived hematopoietic transplants. In this study, nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice were injected with human CD34(+) peripheral blood cells (PBCs) or cord blood cells (CBCs, either enriched for CD34 or density-gradient separated mononuclear cells). Flow cytometric analysis 5 to 11 weeks after transplantation revealed the presence of a human lymphomyeloid hematopoiesis within the murine bone marrow. Immunohistochemical staining of bone marrow cell suspensions using human-specific antibodies showed human cells staining positive for human fibroblast markers, human von Willebrand factor (vWF) and human KDR (vascular endothelial growth factor receptor-2) in mice transplanted with CD34(+) PBCs or CBCs, with mean frequencies between 0.6% and 2.4%. In stromal layers of bone marrow cultures established from the mice, immunohistochemical staining using human-specific antibodies revealed flattened reticular cells or spindle-shaped cells staining positive with human-specific antifibroblast antibodies (mean frequency, 2.2%). Cell populations of more rounded cells stained positive with human-specific antibodies recognizing CD34 (1.5%), vWF (2.2%), and KDR (1.6%). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and subsequent complementary DNA sequencing detected transcripts of human KDR (endothelial specific) and human proline hydroxylase-alpha (fibroblast specific) within the bone marrow and spleen of transplanted mice. Analysis of nontransplanted control mice yielded negative results in immunocytochemistry and RT-PCR. Cells expressing endothelial and fibroblast markers were also detected in the grafts before transplantation, and their numbers increased up to 3 log in vivo after transplantation. These results indicate that stromal progenitor cells are present in human cytokine-mobilized peripheral blood or cord blood that engraft in NOD/SCID mice. (Blood. 2000;96:3971-3978)
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshAntigens, Surface
dc.subject.meshBone Marrow Cells
dc.subject.meshCell Culture Techniques
dc.subject.meshCell Differentiation
dc.subject.meshEndothelium
dc.subject.meshFibroblasts
dc.subject.meshGraft Survival
dc.subject.meshHematopoiesis
dc.subject.meshHematopoietic Stem Cell Transplantation
dc.subject.meshHumans
dc.subject.meshImmunohistochemistry
dc.subject.meshImmunophenotyping
dc.subject.meshMice
dc.subject.meshMice, Inbred NOD
dc.subject.meshMice, SCID
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction
dc.subject.meshStromal Cells
dc.subject.meshTransplantation, Heterologous
dc.titleDonor stromal cells from human blood engraft in NOD/SCID mice.en
dc.typeArticleen
dc.contributor.departmentMax Delbrück Center for Molecular Medicine, Berlin, Germany.en
dc.identifier.journalBlooden
html.description.abstractLittle is known about the presence, frequency, and in vivo proliferative potential of stromal cells within blood-derived hematopoietic transplants. In this study, nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice were injected with human CD34(+) peripheral blood cells (PBCs) or cord blood cells (CBCs, either enriched for CD34 or density-gradient separated mononuclear cells). Flow cytometric analysis 5 to 11 weeks after transplantation revealed the presence of a human lymphomyeloid hematopoiesis within the murine bone marrow. Immunohistochemical staining of bone marrow cell suspensions using human-specific antibodies showed human cells staining positive for human fibroblast markers, human von Willebrand factor (vWF) and human KDR (vascular endothelial growth factor receptor-2) in mice transplanted with CD34(+) PBCs or CBCs, with mean frequencies between 0.6% and 2.4%. In stromal layers of bone marrow cultures established from the mice, immunohistochemical staining using human-specific antibodies revealed flattened reticular cells or spindle-shaped cells staining positive with human-specific antifibroblast antibodies (mean frequency, 2.2%). Cell populations of more rounded cells stained positive with human-specific antibodies recognizing CD34 (1.5%), vWF (2.2%), and KDR (1.6%). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and subsequent complementary DNA sequencing detected transcripts of human KDR (endothelial specific) and human proline hydroxylase-alpha (fibroblast specific) within the bone marrow and spleen of transplanted mice. Analysis of nontransplanted control mice yielded negative results in immunocytochemistry and RT-PCR. Cells expressing endothelial and fibroblast markers were also detected in the grafts before transplantation, and their numbers increased up to 3 log in vivo after transplantation. These results indicate that stromal progenitor cells are present in human cytokine-mobilized peripheral blood or cord blood that engraft in NOD/SCID mice. (Blood. 2000;96:3971-3978)


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