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Ghiasi M.,The Academic Center for Education | Qomi R.T.,The Academic Center for Education | Nikbakht M.,Tehran University of Medical Sciences | Sheykhhasan M.,The Academic Center for Education
Tehran University Medical Journal | Year: 2015

Background: Stem cells represent an ideal cell source for application in tissue engineering and regenerative medicine due to their ability to proliferate and differentiate to a wide variety of cell lineages. With recent development of medical sciences and tissue engineering, usage of adipose-derived mesenchymal stem cells, their culture and differentiation on suitable scaffolds are considered as a successful clinical and research strategy. One of the most crucial factors in a successful tissue engineering technique is to choose an appropriate scaffold which allow cell migration transferring of bioactive factors as well as providing optimal growth environment for stem cells. In this study, the ability of two scaffolds is investigated as a suitable environment for the proliferation and differentiation of adipose-derived mesenchymal stem cells. Methods: This is an in vitro study that was performed in Laboratory of Stem Cell in Academic Center for Education, Culture and Research, Qom province from April 2013 to February 2014. In this study, two scaffolds including fibrin glue and alginate were prepared as two separate groups and after isolating mesenchymal stem cells from adipose tissue and adequate proliferation, they were seeded into each scaffold in chondrogenic medium. After 14 days, the evaluation of viability and gene expression of collagen II and I, SOX9 and aggrecan were done by MTT (3-{4,5-dimethylthiazol-2yl}-2,5diphenyl-2H tetrazolium bromide) assay and real-time PCR technique respectively. Also, cartilaginous tissue formation on scaffolds was evaluated by histological analysis. Results: According to the obtained results, the fibrin glue scaffold showed significant difference in terms of viability in comparison to alginate scaffold in chondrocyte differentiating medium (P< 0.05). Also the results of real-time PCR analysis showed that the fibrin glue scaffold express cartilage specific genes at a higher level than alginate scaffold. Conclusion: The use of natural fibrin glue scaffold can be considered as a suitable en vironment for proliferation and differentiation of adipose-derived mesenchymal stem cells in cartilage tissue engineering. © Tehran University of Medical Sciences. All rights reserved.


Sheykhhasan M.,The Academic Center for Education | Qomi R.T.,The Academic Center for Education | Ghiasi M.,The Academic Center for Education
International Journal of Stem Cells | Year: 2015

Background and Objectives: One of the most cellular source used for cartilage tissue engineering are mesenchymal stem cells (MSCs). In present study, human MSCs were used as cellular source. Since scaffold plays an important role in tissue engineering the aim of this study is to assess fibrin scaffold ability in chondrogenic differentiation of adipose-derived mesenchymal stem cells (ADMSCs). Methods: ADMSCs were isolated and cultured in DMEM medium supplemented with 10% FBS. Also ADMSCs expanded and characterised by flow cytometry. ADMSCs expressed CD44, CD90, CD105 but not CD34. After trypsinization, cells were entered within the fibrin scaffold. Then, chondrogenic medium was added to the scaffold. Seven days after cell culture, cell viability and proliferation were assessed by MTT test. Finally, 14 days after the ending of chondrogenic differentiation, analysis of chondrogenic genes expression was evaluated by RT-PCR and Real time PCR. Also, formation and development of chondrocyte cells was analysed by histological and immunohistochemistry evaluations. Results: Viability and proliferation as well as chondrogenic genes expression within fibrin scaffold increased significantly compared with control group (cells free scaffold). Also, histological and immunohistochemistry evaluation showed that chondrocyte cells and collagen type II are formed on fibrin scaffold. Conclusions: Fibrin is a suitable scaffold for chondrogenic differentiation of ADMSCs. © 2015 Korean Society for Stem Cell Research.


Ghiasi M.,The Academic Center for Education | Kalhor N.,The Academic Center for Education | Tabatabaei Qomi R.,The Academic Center for Education | Sheykhhasan M.,The Academic Center for Education
Frontiers in Life Science | Year: 2015

This study presents a comparative assessment of adipose-derived stem cell (ADSCs) proliferation rates and their viability on five different scaffolds. Five different biomaterial scaffolds were prepared: alginate, poly lactic-co-glycolic acid, fibrin glue, inactive platelet-rich plasma, and active platelet-rich plasma (APRP). Stem cells were isolated from human adipose tissue. Flow cytometry analysis was performed. Specifically, adipogenesis/osteogenesis/chondrogenesis-associated genes expression was analyzed by real-time polymerase chain reaction. These cells were seeded in the prepared scaffolds. After 14 days, the proliferation and viability of MSCs were evaluated using an MTT assay. Also, stemness genes expression was analyzed with the reverse transcriptasepolymerase chain reaction (RT-PCR) method. In addition, the DNA content assay was also performed. The obtained results showed a significant difference between cell proliferation and viability of different scaffolds. APRP and alginate were shown to be the most and least suitable scaffolds in terms of enhancing cell proliferation and maintaining cell viability respectively (p < .05). RT-PCR reactions demonstrated the expression of the various stemness-related markers (Nanog, Octamer4A, and Sox2) when ADSC cells were grown separately on the five different scaffolds. Our study indicates that compared with the scaffolds, APRP could be the best scaffold for support of ADSC proliferation. © 2015 Taylor & Francis


Qomi R.T.,The Academic Center for Education | Sheykhhasan M.,The Academic Center for Education | Kalhor N.,The Academic Center for Education | Ghiasi M.,The Academic Center for Education
Journal of Mazandaran University of Medical Sciences | Year: 2015

Background and purpose: Mesenchymal stem cells (MSCs) are the most widely used cell sources for cartilage tissue engineering. In the present study, human stem cells were used as a cell source. Scaffolds play an important role in tissue engineering, therefore, this study aimed at evaluating the ability of fibrin scaffolds in chondrogenic differentiation of adipose-derived mesenchymal stem cells (ADMSCs). Materials and methods: In this study, after preparing fibrin scaffold, isolating mesenchymal stem cells from adipose tissue, and confirming their isolation the cells were cultured into fibrin scaffolds. Seven days after cell culture, their potential of survival and growth were evaluated using MTT assay. Finally, 14 days after the end of the chondrogenic differentiation, gene expression analysis and chondrocyte morphology formation was performed by Real time PCR and histology analysis, respectively. Results: The mesenchymal stem cell viability, proliferation and expression of 4 specific chondrogenic genes significantly increased in a fibrin scaffold compared with those of the control group. Moreover, the formation of chondrocyte cells on fibrin scaffold was confirmed. Conclusion: Fibrin scaffold was found suitable for condrogenic differentiation of adiposederived mesenchymal stem cells. © 2015, Mazandaran University of Medical Sciences. All rights reserved.

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