Subject(s)
Facility Design and Construction , Laboratories, Hospital , Stem Cell Research , Accreditation/legislation & jurisprudence , Accreditation/standards , Cell Transplantation/legislation & jurisprudence , Clinical Trials as Topic/standards , European Union , Facility Design and Construction/legislation & jurisprudence , Facility Design and Construction/standards , Laboratories, Hospital/legislation & jurisprudence , Laboratories, Hospital/organization & administration , Laboratories, Hospital/standards , Personnel Management/standards , Quality Control , Spain , Stem Cell Research/legislation & jurisprudence , Validation Studies as TopicABSTRACT
BACKGROUND AIMS: The aim of this study was to compare prospectively the vasculogenic capacity of two cell sources, monocytes and CD133+ cells. METHODS: Cells were obtained from healthy donors by adherence or magnetic selection. Animals studies were performed in a model of hind limb ischemia and different groups were established according to type and number of cells infused. Revascularization was measured by sequential blood flow analysis using a laser Doppler device and by assessing capillary density in the ischemic muscles. In order to locate the infused cells, immunofluorescence and immunocytochemistry techniques were performed and analyzed by light and confocal microscopy. RESULTS: During the study period there was a significant improvement in both limb perfusion and capillary density in mice treated with either human monocytes or CD133+ cells (P<0.05) compared with non-treated mice. No cells were detected as incorporated into the vessels when 1 x 10(5) cells were used but with higher doses (1 x 10(6)) a few human cells were observed integrated into the vessels in both groups of treated mice. Supernatants of both cell types showed vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and platelet-derived growth factor- AB (PDGF-AB) expression. CONCLUSIONS: Treatment with human monocytes or CD133+ cells improves blood perfusion and capillary density in a murine model and both cell types seem to stimulate vasculogenesis in a fairly similar way.
Subject(s)
Antigens, CD/metabolism , Glycoproteins/metabolism , Hindlimb/blood supply , Hindlimb/pathology , Ischemia/pathology , Monocytes/cytology , Neovascularization, Physiologic , Peptides/metabolism , AC133 Antigen , Angiogenesis Inducing Agents/metabolism , Animals , Capillaries/pathology , Cell Movement , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Humans , Immunophenotyping , Laser-Doppler Flowmetry , Mice , Microscopy, Confocal , Muscles/pathology , Perfusion , Phenotype , Regional Blood Flow , Staining and LabelingABSTRACT
BACKGROUND AND OBJECTIVES: Whether human mesenchymal stem cells (MSC) can be transplanted is controversial and their presence in peripheral blood is not fully accepted. In the present study we have analyzed whether, within the allogeneic transplantation setting, MSC are of host or donor origin. DESIGN AND METHODS: Bone marrow MSC from 19 patients who had undergone allogeneic transplantation were expanded and identified using immunophenotypic markers. After that, chimerism studies were performed using reverse transcription polymerase chain reaction of short tandem repeat (STR) loci. Analyses were carried out at different time-points after transplantation, with a total of 44 samples studied. Bone marrow was used as the source of stem cells for transplantation in 4 cases and peripheral blood in 15 cases. The conditioning regimen was standard in 9 patients and non-myeloablative in 10 patients. RESULTS: Our results show that in the great majority of cases analyzed (17 out 19), MSC were of host origin. However, in 2 patients with multiple myeloma who had received a reduced intensity transplantation using peripheral blood stem cells, MSC were partially of donor origin (60.17% and 26.13% of total MSC). INTERPRETATION AND CONCLUSIONS: These findings indicate that after allogeneic transplantation MSC from the donor can engraft in bone marrow. Moreover, since the stem cells were obtained from peripheral blood, it can be concluded that MSC circulate among mobilized peripheral blood stem cells and can engraft in bone marrow after allogeneic transplantation.
