Pigliaru, Giovanni (2015) Studi molecolari sulla plasticità e differenziamento di cellule mesenchimali staminali durante l’invecchiamento in vitro. Doctoral Thesis.
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Aim: Recent evidence suggests that ageing-related diseases could result in an accelerated loss of self- renewal capability of adult stem cells, normally involved in replacing damaged cellular elements. In previous works, we highlighted that a specific treatment, named tissue optimization-regenerative (TO-RGN), of radio- electric asymmetric conveyer (REAC) technology, influenced gene expression profiles controlling stem cell differentiation and pluripotency of human skin-derived fibroblasts in vitro. The purpose of the present work was to verify whether TO-RGN may also be effective in counteracting the expression of the senescence marker beta-galactosidase and of senescence-associated gene expression patterning, engaged during prolonged culture of human adipose-derived stem cells (hADSCs). We analyzed the Characterization of hADSCs by flow cytometry Analysis, Adipogenesis, osteogenesis, and vasculogenesis in culture, Gene expression, Immunoblotting analysis and the assessment of telomere length.
Methods: According to the policy approved by the local ethical committee of the University of Bologna (title of approved project: “Assessment of mesenchymal stem cells in human adipose tissue,” code number 013/2010/O/Tess, date of approval from the ethical committee: 16 February 2010), all tissue samples were obtained after informed consent. All participants, according to the consent procedure approved by the ethical committee, provided their written consent specifically dedicated to the harvesting of their abdominal fat (50 – 100 mL of lipoaspirate).
Results: TO-RGN treatment remarkably influenced cell viability, as shown by its ability to significantly enhance the number of vital cells as compared to untreated controls. SA- β –Gal staining in hADSCs cultured in the absence or presence of TO-RGN throughout an observation period of 90 days. In the present work, we analyzed the effect of TO-RGN treatment on hADSC senescence in vitro, and demonstrated that this physical treatment, without the aid of viral vectors, was able to counteract biochemical and morphological changes occurring in stem cells during aging.
Conclusion: In the present work, we analysed the effect of TO-RGN treatment on hADSC senescence in vitro, and demonstrated that this physical treatment, without the aid of viral vectors, was able to counteract biochemical and morphological changes occurring in stem cells during aging.
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