The Effect of BMP-6 Growth Factor on Differentiation of Adipose-derived Stem Cells into Chondrocyte in Pellet Culture System

Document Type : Original Article(s)

Authors

1 Assistant Professor, Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan

2 Associate Professor, Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan

3 Professor, Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan

4 Associate Professor, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

5 Assistant Professor, Department of Physics and Medical Engineering, School of Medicine, Isfahan University of Medical Sciences, Isfahan

6 Assistant Professor, Department of Pathology and Hematology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

7 Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan

8 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

Abstract

Background:Cartilage damaged by trauma or degenerative arthritis has a limited capacity for regeneration most likely because of its avascularity and low cellularity. In this study, adipose–derived stem cells were cultured in chondrogenic medium supplemented isolated from adipose tissue then were induced for chondrogenesis with BMP-6 in pellet culture system.

Methods: Human adipose tissues obtained from cesarean section and ADSCs were isolated and characterized by flowcytometry techniques for CD14, CD44, CD45, and CD90 markers. To induct of chondrogenesis ADSCs were cultured in pellet culture for 21 days in chondrogenic medium supplemented with BMP-6. Resulted constructs were investigated by histological, immunohistochemical and RT-PCR methods.

Findingss: ADSCs were expressed CD44, CD90 and not CD14 or CD45. ADSCs chondrogenesis was occurred successfully. Stained-sections in H-E and alcian blue were showed chondrocytes in lacuna with sulfated proteoglycans extracellular matrix. Existence of collagen type-II and aggrecan was proved in constructs by immunohistochemical study. RT-PCR method was exhibited collagen type-II and aggrecan genes expression in differentiated cells.

Conclusion: Human adipose-derived stem cells are excellent candidate for cartilage tissue engineering. In pellet culture ADSCs in presence of BMP-6 were induced for chondrogenesis.

Key word: Chondrogenesis, Adipose-derived stem cells, Tissue engineering, Collagen type II, BMP-6, Pellet culture system.