RESUMEN
It is well-known that the modified gravity affects not only the organism as a whole, but also the molecular processes in individual cells. The goal of this study was to investigate the influence of simiulated microgravity (SMG) on gene expression pattern in cultured human bone marrow mesenchymal stem cells (hMSCs) after prolonged exposure to SMG on dRPM. It was found that the expression level of 144 genes significantly changed in hMSCs after 20 days of exposure to SMG: the expression of 30 genes significantly increased and 114 - decreased. Gene analysis revealed that these genes can be attributed to the 11 major categories corresponding to its biological role in the cells. The greatest number of genes with altered expression was found to be in the groups "Matrix and Adhesion", "Inflammation and Cell-to-Cell Communications", "Metabolism" and "Signalling". Interestingly, some of the genes with down-regulated expression were related to the bone development and differentiation (COL15A1, CXCL12, DPTand WISP2) or participated in the cell-to-cell communications ofhMSCs with other cell types (CXCL12, SDF4). The obtained results indicate that the changes in gene expression in bone marrow progenitor cells can be involved into space flight-induced osteopenia.
Asunto(s)
Enfermedades Óseas Metabólicas/metabolismo , Células de la Médula Ósea/metabolismo , Diferenciación Celular , Regulación de la Expresión Génica , Células Madre Mesenquimatosas/metabolismo , Osteogénesis , Ingravidez/efectos adversos , Enfermedades Óseas Metabólicas/etiología , Enfermedades Óseas Metabólicas/patología , Células de la Médula Ósea/patología , Células Cultivadas , Humanos , Células Madre Mesenquimatosas/patologíaRESUMEN
Morphofunctional state and differential potential of cultured osteoblasts and osteogenic precursor's cells under real and simulated microgravity are in the focus of presented review. Summary of data of osteoblast cell responses to altered microgravitational circumstances are discussed. Mesenchymal stromal cells are the population of multipotential precursors of adult bone marrow stroma able to differentiate into osteogenic. Adipogenic and chondrogenic lineages. Recent studies have demonstrated that multipotential mesenchymal stromal cells can be the real candidates to gravity-responding cell type and can be involved into space flight-induced osteopenia. These findings determine the necessity of more detailed study of its biology in vitro and in vivo.
Asunto(s)
Adaptación Fisiológica , Enfermedades Óseas Metabólicas/metabolismo , Células de la Médula Ósea/metabolismo , Células Madre Mesenquimatosas/metabolismo , Células Madre Multipotentes/metabolismo , Células del Estroma/metabolismo , Ingravidez/efectos adversos , Adipocitos/metabolismo , Adipogénesis , Adulto , Animales , Enfermedades Óseas Metabólicas/etiología , Condrocitos/metabolismo , Humanos , Osteogénesis , Vuelo EspacialRESUMEN
Effects of simulated microgravity (cell clinostatting and containment in the Random Positioning Machine (RPM), Dutch Space, The Netherlands) on interleukins production by mesenchymal stromal cells (MSC) of the human marrow and MSC osteogenous derivatives obtained through cell stimulation by growth factors (10(-8) M dexamethazone, 0.2 mM ascorbic acid, 10 mM beta-glycerol-phosphate) were studied. Twenty-day clinostatting was found to increase 1.4 to 3.2 times the interleukin-8 (IL-8) content in the MSC and MSC osteogenous derivatives culture medium. Microgravity effects simulated with the use of RPM raised the IL-8 production 1.5 - 6 times and 1.6-2.1 times on the average after 10 days and 20 days of containment, respectively. MSC and MSC osteogenous derivatives demonstrated a downward trend in IL-6 secretion in the culture put in RPM. Therefore, simulation of the microgravity effects using different systems modifies interleukins production by MSC and also by mature cells of the osteoblastic phenotype.
Asunto(s)
Interleucina-6/biosíntesis , Interleucina-8/biosíntesis , Células Madre Mesenquimatosas/metabolismo , Osteogénesis/fisiología , Simulación de Ingravidez , Células Cultivadas , Estudios de Seguimiento , Humanos , Células Madre Mesenquimatosas/citologíaRESUMEN
Cytoskeletal alterations occur is several cell types including lymphocytes, glial cells, and osteoblasts, during spaceflight and under simulated microgravity (SMG). One of the potential mechanisms of cytoskeletal gravisensitivity of a cell is disruption of extracellular matrix and integrin interactions. Therefore, we investigated the effects of SMG on F-actin cytoskeleton structure, vinculin focal adhesions, expression of some integrin subtypes and cellular adhesion molecules in mesenchymal stem cells (hMSCs) derived from human bone marrow in vitro. Simulated microgravity was produced by RPM (manufactured by Dutch Space, The Netherlands). Culture flasks with MSCs were settled on the inner platform of RPM for 30 minutes, 6, 24, 48 and 120 h. The results have shown that actin cytoskeleton is very fast reorganized even after 30 minutes of simulated microgravity. The number of cells with disrupted actin cytoskeleton steadily increased with the increasing exposure time. However these changes were reversible because the cells partly recovered their F-actin structure after 120-hours-exposure. In addition, we observed vinculin redistribution inside the cells after 6 hours and subsequent terms of exposures. This process was accompanied with increasing of fluorescence intensity of cells. The expression of integrin-alpha2 increased after exposure in RPM. Also we observed decrease in the number of VCAM-1-positive cells and changes in the expression of ICAM-1. Thus, our findings indicate that SMG leads to reversible microfilament alterations of hMSCs and alters adhesion properties of this type of cells.
Asunto(s)
Citoesqueleto/ultraestructura , Células Madre Mesenquimatosas/ultraestructura , Ingravidez , Actinas/metabolismo , Adhesión Celular , Citoesqueleto/metabolismo , Humanos , Integrina alfa2/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/fisiología , Células del Estroma/metabolismo , Células del Estroma/fisiología , Células del Estroma/ultraestructura , Factores de Tiempo , Molécula 1 de Adhesión Celular Vascular/metabolismo , Vinculina/metabolismoRESUMEN
The effect of immunization with the synthetic fragments of the alpha7 subunit of the acetylcholine nicotine receptor on the spatial memory of mice subjected to olfactory bulbectomy, which causes the development of the neuro-degenetrative disease of Alzheimer's type, was studied. Mice of the NMRI line were immunized with the KLH conjugates of two peptide fragments of the N-terminal fragment of the alpha7 subunit extraxcellular fragment, subjected to olfactory bulbectomy to cause the development of the neurodegenetrative disease of Alzheimer's type, and then the state of the spartial memory was evaluated. It was shown that 20% of bulbectomized mice immunized with the N-terminal 1-23 fragment exhibited good spatial memory after training. Immunization with the peptide construct (159-167)-(179-188) consisting of two hydrophilic exposed regions of alpha7-subunit induced good spatial memory in 50% of bulbectomized mice, while in the control group, which received only KLH, none of the animals were educated. Thus, the development of immunotherapy with peptide (159-167)-(179-188) seems to be a promising approach to prophylaxis and treatment of Alzheimer's disease.