RESUMEN
BACKGROUND: Mesenchymal stromal cells (MSCs) are multipotent progenitor cells used in several cell therapies. MSCs are characterized by the expression of CD73, CD90, and CD105 cell markers, and the absence of CD34, CD45, CD11a, CD19, and HLA-DR cell markers. CD90 is a glycoprotein present in the MSC membranes and also in adult cells and cancer stem cells. The role of CD90 in MSCs remains unknown. Here, we sought to analyse the role that CD90 plays in the characteristic properties of in vitro expanded human MSCs. METHODS: We investigated the function of CD90 with regard to morphology, proliferation rate, suppression of T-cell proliferation, and osteogenic/adipogenic differentiation of MSCs by reducing the expression of this marker using CD90-target small hairpin RNA lentiviral vectors. RESULTS: The present study shows that a reduction in CD90 expression enhances the osteogenic and adipogenic differentiation of MSCs in vitro and, unexpectedly, causes a decrease in CD44 and CD166 expression. CONCLUSION: Our study suggests that CD90 controls the differentiation of MSCs by acting as an obstacle in the pathway of differentiation commitment. This may be overcome in the presence of the correct differentiation stimuli, supporting the idea that CD90 level manipulation may lead to more efficient differentiation rates in vitro.
Asunto(s)
Adipocitos/metabolismo , Silenciador del Gen , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/metabolismo , Antígenos Thy-1/genética , Adipocitos/citología , Tejido Adiposo/citología , Tejido Adiposo/metabolismo , Líquido Amniótico/citología , Líquido Amniótico/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Moléculas de Adhesión Celular Neuronal/genética , Moléculas de Adhesión Celular Neuronal/metabolismo , Diferenciación Celular , Proliferación Celular , Pulpa Dental/citología , Pulpa Dental/metabolismo , Proteínas Fetales/genética , Proteínas Fetales/metabolismo , Humanos , Receptores de Hialuranos/genética , Receptores de Hialuranos/metabolismo , Inmunofenotipificación , Lentivirus/genética , Lentivirus/metabolismo , Células Madre Mesenquimatosas/citología , Osteoblastos/citología , Cultivo Primario de Células , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Linfocitos T/citología , Linfocitos T/metabolismo , Antígenos Thy-1/metabolismoRESUMEN
Here we describe multimodal iron oxide nanoparticles conjugated to Rhodamine-B (MION-Rh), their stability in culture medium, and subsequent validation of an in vitro protocol to label mesenchymal stem cells from umbilical cord blood (UC-MSC) with MION-Rh. These cells showed robust labeling in vitro without impairment of their functional properties, the viability of which were evaluated by proliferation kinetic and ultrastructural analyzes. Thus, labeled cells were infused into striatum of adult male rats of animal model that mimic late onset of Parkinson's disease and, after 15 days, it was observed that cells migrated along the medial forebrain bundle to the substantia nigra as hypointense spots in T2 magnetic resonance imaging. These data were supported by short-term magnetic resonance imaging. Studies were performed in vivo, which showed that about 5 × 10(5) cells could be efficiently detected in the short term following infusion. Our results indicate that these labeled cells can be efficiently tracked in a neurodegenerative disease model.
Asunto(s)
Rastreo Celular/métodos , Sangre Fetal/citología , Nanopartículas de Magnetita , Células Madre Mesenquimatosas/citología , Animales , Diferenciación Celular , Movimiento Celular , Trasplante de Células Madre de Sangre del Cordón Umbilical , Femenino , Colorantes Fluorescentes , Humanos , Imagen por Resonancia Magnética , Nanopartículas de Magnetita/química , Masculino , Trasplante de Células Madre Mesenquimatosas , Nanomedicina , Enfermedades Neurodegenerativas/patología , Enfermedades Neurodegenerativas/terapia , Embarazo , Ratas , Ratas Wistar , Rodaminas , Sustancia Negra/citologíaRESUMEN
BACKGROUND: Nanoparticles in suspension are often utilized for intracellular labeling and evaluation of toxicity in experiments conducted in vitro. The purpose of this study was to undertake a computational modeling analysis of the deposition kinetics of a magnetite nanoparticle agglomerate in cell culture medium. METHODS: Finite difference methods and the Crank-Nicolson algorithm were used to solve the equation of mass transport in order to analyze concentration profiles and dose deposition. Theoretical data were confirmed by experimental magnetic resonance imaging. RESULTS: Different behavior in the dose fraction deposited was found for magnetic nanoparticles up to 50 nm in diameter when compared with magnetic nanoparticles of a larger diameter. Small changes in the dispersion factor cause variations of up to 22% in the dose deposited. The experimental data confirmed the theoretical results. CONCLUSION: These findings are important in planning for nanomaterial absorption, because they provide valuable information for efficient intracellular labeling and control toxicity. This model enables determination of the in vitro transport behavior of specific magnetic nanoparticles, which is also relevant to other models that use cellular components and particle absorption processes.
Asunto(s)
Nanopartículas de Magnetita/química , Modelos Teóricos , Algoritmos , Simulación por Computador , Convección , Medios de Cultivo/química , Difusión , Cinética , Tamaño de la Partícula , Suspensiones/químicaRESUMEN
Gliomas are a group of heterogeneous primary central nervous system (CNS) tumors arising from the glial cells. Malignant gliomas account for a majority of malignant primary CNS tumors and are associated with high morbidity and mortality. Glioblastoma is the most frequent and malignant glioma, and despite the recent advances in diagnosis and new treatment options, its prognosis remains dismal. New opportunities for the development of effective therapies for malignant gliomas are urgently needed. Magnetic hyperthermia (MHT), which consists of heat generation in the region of the tumor through the application of magnetic nanoparticles subjected to an alternating magnetic field (AMF), has shown positive results in both preclinical and clinical assays. The aim of this review is to assess the relevance of hyperthermia induced by magnetic nanoparticles in the treatment of gliomas and to note the possible variations of the technique and its implication on the effectiveness of the treatment. We performed an electronic search in the literature from January 1990 to October 2010, in various databases, and after application of the inclusion criteria we obtained a total of 15 articles. In vitro studies and studies using animal models showed that MHT was effective in the promotion of tumor cell death and reduction of tumor mass or increase in survival. Two clinical studies showed that MHT could be applied safely and with few side effects. Some studies suggested that mechanisms of cell death, such as apoptosis, necrosis, and antitumor immune response were triggered by MHT. Based on these data, we could conclude that MHT proved to be efficient in most of the experiments, and that the improvement of the nanocomposites as well as the AMF equipment might contribute toward establishing MHT as a promising tool in the treatment of malignant gliomas.
Asunto(s)
Glioma/terapia , Hipertermia Inducida/métodos , Nanopartículas de Magnetita/uso terapéutico , Animales , HumanosRESUMEN
Adherent umbilical cord blood stromal cells (AUCBSCs) are multipotent cells with differentiation capacities. Therefore, these cells have been investigated for their potential in cell-based therapies. Quantum Dots (QDs) are an alternative to organic dyes and fluorescent proteins because of their long-term photostability. In this study we determined the effects of the cell passage on AUCBSCs morphology, phenotype, and differentiation potential. QDs labeled AUCBSCs in the fourth cell passage were differentiated in the three mesodermal lineages and were evaluated using cytochemical methods and transmission electron microscopy (TEM). Gene and protein expression of the AUCBSCs immunophenotypic markers were also evaluated in the labeled cells by real-time quantitative PCR and flow cytometry. In this study we were able to define the best cellular passage to work with AUCBSCs and we also demonstrated that the use of fluorescent QDs can be an efficient nano-biotechnological tool in differentiation studies because labeled cells do not have their characteristics compromised.
Asunto(s)
Adhesión Celular , Diferenciación Celular , Proliferación Celular , Sangre Fetal/citología , Nanotecnología , Células del Estroma/citología , Recuento de Células , Linaje de la Célula , Células Cultivadas , Sangre Fetal/metabolismo , Citometría de Flujo , Humanos , Inmunofenotipificación , Células del Estroma/metabolismo , Células del Estroma/ultraestructuraRESUMEN
The aim of the present work is the presentation of a quantification methodology for the control of the amount of superparamagnetic iron oxide nanoparticles (SPIONs) administered in biological materials by means of the ferromagnetic resonance technique (FMR) applied to studies both in vivo and in vitro. The in vivo study consisted in the analysis of the elimination and biodistribution kinetics of SPIONs after intravenous administration in Wistar rats. The results were corroborated by X-ray fluorescence. For the in vitro study, a quantitative analysis of the concentration of SPIONs bound to the specific AC133 monoclonal antibodies was carried out in order to detect the expression of the antigenic epitopes (CD133) in stem cells from human umbilical cord blood. In both studies FMR has proven to be an efficient technique for the SPIONs quantification per volume unit (in vivo) or per labeled cell (in vitro).