Evaluation Of DNA Damage Durig Differentiation Of Mesenchymal Stem Cells To Germ-like Cells Based On Retinoic Acid

Salem, Maryam and Bayrami, Abolfazl and Mirzapour, Tooba and Sagha, Mohsen and Asadi, Asadollah (2016) Evaluation Of DNA Damage Durig Differentiation Of Mesenchymal Stem Cells To Germ-like Cells Based On Retinoic Acid. Masters thesis, University of Mohaghegh Ardabili.

Text (بررسی آسیب های DNA در طی تمایز سلول‌های بنیادی مزانشیمی به سلول‌های مشابه جرم تحت تاثیر رتینوئیک اسید) Maryam Salem.pdf Download (1MB)

Abstract

Infertility in men reason of 30%of couple's infertility.Disruption of genetic material is important because it lead to incomplete transmission of hereditary information to fetus.For this reason preserving the integrity and accuracy of DNA in the nuclei of sex cells, for transferring genetic material from one generation to another perfect is very important.In this study, Mesenchymal stem cells were extracted from rat bone marrow aspiration procedure. The cells initially were for 3 days under the influence of different doses of retinoic acid (….mM) and were evaluated by MTT. MTT results showed the viability of the cells in 10-5 and 10-6 M Retinoic acid is more than other doses.Since the differentiation groups were 10 and 15 days. So this time Mesenchymal cells were for 10 days under the influence of 10-5 and 10-6 M of Retinoic acid. So this dose was selected as the dose of differentiation.Then four groups: Control group, induced group with condition medium, induced group with 10-6 M Retinoic acid and induced group with 10-6 M Retinoic acid accompany condition medium that were cultured for 10 and 15 days.The first part of our study showed that the group of condition medium-induced in the expression of premeiotic markers OCT4 and PLZF and meiosis marker SCP3 differentiate induced bone marrow Mesenchymal stem cells intogerm-like cells up to meiotic phase has more effect.In the second part we studied the four groups for DNA damage. MTT studies and Acridine Orange and DAPI staining showed that the treated group with the dose of 106 MRetinoic cause of apoptosis in the cells at the differentiation day of ten and on the fifteenth day of differentiation significant differences was observed in apoptosis of treated cells compared to control group.Conditioned Medium-induced cells showed apoptosis less than other groups.Fragmentation was not observed in any of the groups. So apoptosis in the group that treated with 10-6 M Retinoic acid not enough to cause DNA fragmentation. Finally all groups were examined for the expression of apoptotic genes index. The apoptosis was observed in treated cells with condition medium, Retinoic acid, Retinoic acid accompany condition medium. Probably another apoptotic factors incondition medium which raises the level of gene expression. Eventually the result showed that condition medium treated group both in terms of expression of differentiation markers and low rate of apoptosis is more suitable than other groups.

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