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dc.contributor Amleh, Asma
dc.contributor.author Alaasar, Walid Almoafy
dc.date.accessioned 2017-09-14T09:19:05Z
dc.date.created 2017-09-11
dc.date.issued 2017-09-14
dc.identifier.uri http://dar.aucegypt.edu/handle/10526/5202
dc.description Mouse fibroblasts L929 were used for this analysis, study cells were incubated with osteogenic differentiation media containing Dexamethazone, Ascorbic acid and Beta Glycerol phosphate for 21 days during which the media were changed every 3 days, Alizarin red stain was applied to monitor the deposition of calcium in the induced and un-induced L929 cells after 2 and 3 weeks from the onset of induction. The same timetable has been followed by taking aliquots of control and study cells and processing them for RNA extraction followed by cDNA synthesis and PCR amplification using primers for specific osteoblasts genes (Runx2, Osteocalcin and Collagen1) ( Ericet al., 2010), Cobra1 gene primers and primers of B-actin as a control. The products of RT-PCR were run on a gel, and transcription analysis was done. en_US
dc.description.abstract Fibroblasts are the most common connective tissue cells and are responsible for the synthesis of extracellular matrix (ECM) and collagen. They are mainly of mesenchymal stem cells (MSCs) origin. Fibroblasts are fully differentiated cells although some recent findings reported that fibroblasts possess some potency that allows them to differentiate into other cells types. They are said to undergo adipogenic, chondrogenic, hepatogenic, nurogenic and osteogenic differentiation if grown in the suitable conditions. In the normal physiologic environment, fibroblasts can also differentiate to epithelial cells, a phenomenon known as the mesenchymal epithelial transition (MET). In the current research, we induced L929 mouse fibroblasts to differentiate into osteoblasts cells using the growth media containing Dexamethazone, Ascorbic acid and Beta-Glycerol phosphate which are the substances known to promote osteogenic differentiation. We have increased the concentration of dexamethazone than that used previously, and investigated the effect of this increase on the time of induction. We also investigated the levels of Cobra1 (which is an integral member of the negative elongation (NELF) complex) mRNA in L929 that underwent osteogenic induction compared to the non-induced ones. We show a successful induction the onset of which is earlier than previously reported in the literature. Furthermore, to the best our knowledge our current data, for the first time, suggest a potential role of Cobra1 in maintaining the plasticity of L929. en_US
dc.description.sponsorship I would like to thank Dr. Asma Amleh for her support and patience. I am also grateful for Dr. Adham Ramadan's support. Great help was offered to me from my colleagues; Noha Saad, Nancy Hasanein, Myret Ghabriel, Menna Ghoraba and Menna Alfar. So thank them all so much. Finally, I would like to express much gratitude to the AUC for providing the grant that funded this research. en_US
dc.format.medium theses en_US
dc.language.iso en en_US
dc.rights Author retains all rights with regard to copyright. en
dc.subject Direct conversion of cells fate en_US
dc.subject COBRA1 expression en_US
dc.title Direct conversion of mouse fibroblasts into osteoblasts and investigating COBRA1 expression responce en_US
dc.type Text en_US
dc.subject.discipline Biotechnology en_US
dc.rights.access This item is restricted for 2 years from the date issued en_US
dc.contributor.department American University in Cairo. Dept. of Biology en_US
dc.embargo.lift 2019-09-14T09:19:05Z


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