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Stem cellsOnly 14 pages are availabe for public view
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Abstract
Aging is a universal process that began with the origination of life. Accumulation of the diverse deleterious changes produced by aging throughout the cells and tissues progressively impairs function and can eventually cause death. Mesenchymal stem cells (MSCs) have been used in cell-based therapy for various diseases, due to their immunomodulatory and anti-inflammatory effects. Also considered effective strategy to protect against tissue and organ injury. MSC transplantation also serves as a promising therapy for regenerative medicine However, the function of MSCs is known to decline with age, a process that is called senescence. Senescence in mesenchymal stem cells (MSCs) not only contributes to organism aging and the development of a variety of diseases but also severely impairs their therapeutic properties as a promising cell therapy and MSC-based regenerative medicine. Resveratrol, a small molecule found in red wine, is reported to slow aging in simple eukaryotes and has been suggested as a potential calorie restriction mimetic. Resveratrol has also been reported to act as a sirtuin activator, and this property has been proposed to account for its anti-aging effects. So the aim of our work is to evaluate the possible effects of resveratrol to improve and even retard BM-MSC aging, keep their stemness and improve their proliferative kinetics, decrease aneuploid cells monitor cell mitotic division and maintain genomic stability. A total number of 35 female albino rats were used. Fifteen rats aged 9 months and tweny rats aged fifteen months Animals were divided into 2 main age groups 1-group Ι: 9 month age group: included 15 animals, further subdivided into three subgroups: control group, one week resveratrol treated group and two week resveratrol treated groups (5 rats were included in each group) 2-group ΙΙ: 15 month age group included 20 animals, further subdivided into four subgroups; control group, one week resveratrol treated group ,two week resveratrol treated groups and one month resveratrol treated group (5 rats were included in each group). Resveratrol was regularly administrated daily orally at a dose of 2.5 mg/kg dissolved in 0.5 ml PBS. At the designated times, the animals were sacrificed, BM-MSCS were isolated and cultivated ,monitored ,tracked for sequential periods up to (80-90%) confluence and photographed by inverted microscope for morphological study. Subculture and trypsinization then was performed ; cell pellet was suspended and about 10.000 cells was prepared for flow cytometric analysis, characterization detecting CD90,CD31 and CD34 marker expression and DNA index analysis ; remaining cells were recultivated in 6-well plate for measuring MTT,CFU,PDT and ICC immunocytohemical staining. The results of this study: Resveratrol was found to enhance cellular proliferation by minimizing time to reach full confluent state in duration dependent manner; Regarding BM-MSCS of 9 months age group optimizing resveratrol treatment duration was obtained only after two week while in 15 months age group we could optimize duration of resveratrol treatment to extend up to one month to obtain significant satisfactory results. Cells derived from resveratrol treated animals tended to exhibit mainly spindle shape and syncytial like morphological pattern unlike those derived from control which exhibited various spindle, branched and sometimes hypertrophic pattern. Resveratrol improved CD90 marker expression correlated with duration of resveratrol treatment. Resveratrol could improve proliferation kinetic parameters and aneuploidy cell proportion having significant results even after one week resveratrol treatment in cells derived from 9 months age group but starting from two week resveratrol treatment and up in cells derived from 15 months age group. Resveratrol could improve Bubr1 marker expression in duration dependent manner demonstrated by intensifying immunocytochemical reaction in both age groups.