The Expression of Osteogenic Markers in Differentiated Osteoblasts From Adipose-Derived Stem Cells in Pellet Culture and Monolayer Systems



Heirat B¹ ; Akbarzadeh S¹ ; Sadeghi F^{2, 4} ; Ghorbani M³ ; Bahrambeigi V^{2, 4} ; Goharian V⁵ ; Hashemibani B^{2, 4} Show Affiliations
Source: Journal of Isfahan Medical School Published:2012

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.