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In recent years significant progress has been witnessed in the identification of stem cells, which have now also been identified in the lungs. The aim of this was to induce post-pneumonia alveolar regeneration to facilitate the identification of stem cells. The studies were performed on Buffalo strain rats. Pneumonia was induced in the animals by a sub-pleural injection of carragenin. On days 4, 5 and 10 of the experiment both the control and experimental animals received intraperitoneal injections of bromodeoxyuridine (BrdU). Twenty-four hours after the last BrdU injection the rats were sacrificed and samples of the lungs were taken for examination. In order to detect proliferating cells in the paraffin sections, BrdU incorporation was detected with monoclonal antibodies. In pilot experiments BrdU incorporation was demonstrated in individual alveolar cells of variable distribution and of variable intensity in the colour reaction. The results have confirmed the existence of stem cells in pulmonary alveoli but their closer characterisation requires further studies with other techniques to detect pulmonary stem cells.
The cancer stem cell theory elucidates not only the issue of tumour initiation and development, tumour’s ability to metastasise and reoccur, but also the ineffectiveness of conventional cancer therapy. This review examines stem cell properties, such as self-renewal, heterogeneity, and resistance to apoptosis. The ‘niche’ hypothesis is presented, and mechanisms of division, differentiation, self-renewal and signalling pathway regulation are explained. Epigenetic alterations and mutations of genes responsible for signal transmission may promote the formation of cancer stem cells. We also present the history of development of the cancer stem cell theory and discuss the experiments that led to the discovery and confirmation of the existence of cancer stem cells. Potential clinical applications are also considered, including therapeutic models aimed at selective elimination of cancer stem cells or induction of their proper differentiation.
microRNAs regulate all the cellular processes, and are strongly involved in differentiation of stem cells. Disturbances in the regulation of microRNAs expression and activity may deviate the stem cells fate, impairing their differentiation and contributing to diseases initiation or progression. In this talk the role of microRNAs in certain neurological conditions will be discussed.
Ischemic diseases are characterized by the presence of pro-apoptotic stimuli, which initiate a cascade of processes that lead to cell injury and death. Several molecules and events represent detectable indicators of the different stages of apoptosis. Among these indicators is phosphatidylserine (PS) translocation from the inner to the outer leaflet of the plasma membrane, which can be detected by annexinV (ANXA5) conjugation. This is a widely used in vivo and in vitro assay marking the early stages of apoptosis. We report here on an original method that employs PS-ANXA5 conjugation to target stem cells to apoptotic cells. Mesenchymal stem cells (MSCs) from GFP-positive transgenic rats were biotinylated on membrane surfaces with sulfosuccinimidyl-6-(biotinamido) hexanoate (sulfo-NHS-LC-biot) and then bound to avidin. The avidin-biotinylated MSCs were labeled with biotin conjugated ANXA5. Bovine aortic endothelial cells (BAE-1 cells) were exposed to UVC to induce caspasedependent apoptosis. Finally, we tested the ability of ANXA5-labeled MSCs to bind BAE-1 apoptotic cells: suspended ANXA5-labeled MSCs were seeded for 1 hour on a monolayer of UV-treated or control BAE-1 cells. After washing, the number of MSCs bound to BAE-1 cells was evaluated by confocal microscopy. Statistical analysis demonstrated a significant increase in the number of MSCs tagged to apoptotic BAE-1 cells. Therefore, stem cell ANXA5 tagging via biotin-avidin bridges could be a straightforward method of improving homing to apoptotic tissues.
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