Document Type : Original Article (s)
Authors
1
MSc Student, Department of Biology, School of Basic Sciences, Jahrom Branch, Islamic Azad University, Jahrom, Iran
2
Department of Anatomy and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3
PhD Student, Department of Tissue Engineering, School of Modern Technologies, Tehran University of Medical Sciences, Tehran, Iran
4
MSc Student, Department of Anatomy and Molecular Biology, School of Medicine And Student Research committee, Isfahan University of Medical Sciences, Isfahan, Iran
5
Department of Surgery, Amin Hospital, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
6
Assistant Professor, Department of Anatomy and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Abstract
Background: Nowadays high accident rates, fractures leading to permanent bone disorders, and the impossibility of bone transplant have made scientists to look for new methods of repairing injured bones. Considering the application of stem cells in bone tissue engineering, there exists the necessity to investigate various culture methods and suitable fields and scaffolds. Thus, we decided to induce adipose-derived stem cells into osteoblast cells in two systems of pellet culture and monolayer and compare osteogenic markers. Methods: Stem cells have been separated via mechanical and enzymatic methods and cultured in osteogenic medium. Then, RNA was separated from differentiated cells, complementary DNA (cDNA) was synthesized and amplified. Polymerase chain reaction (PCR) product was transferred to electrophoresis gel. The intensity of the bands was measured by Image-J software and analyzed by SPSS. Findings: Osteocalcin gene expression in differentiated cells in the two culture systems showed no significant difference. Therefore, the expression of osteocalcin gene in both monolayer and pellet systems is the same and acceptable. However, average osteopontin gene in differentiated cells in the two culture systems showed a significant difference. The expression of osteopontin gene in pellet system was more than monolayer systems. Conclusion: This study indicated that pellet and monolayer culture systems are appropriate for bone engineering but osteocalcin and osteopontin gene expressions were different in the two culture system. Keywords: Adipose-derived stem cells, Osteocalcin, Osteopontin, Pellet culture
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