Effects of soft substrates on the chondrogenic differentiation of human synovial-derived mesenchymal stem cells
10.3969/j.issn.2095-4344.2015.50.025
- VernacularTitle:软基质力学特性对滑膜间充质干细胞向软骨细胞分化的干预作用
- Author:
Shuangshuang CUI
;
Zhaozhen YU
;
Shunlu YU
;
Wenjun ZHAO
;
Likun ZHAO
;
Guosheng XING
;
Xiaoyuan DUAN
- Publication Type:Journal Article
- Keywords:
Synovial Membrane;
Mesenchymal Stem Cels;
Acrylamides;
Elastic Modulus;
Transforming Growth Factor beta1;
Chondrocytes;
Cel Differentiation;
Tissue Engineering
- From:
Chinese Journal of Tissue Engineering Research
2015;(50):8177-8183
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Our previous studies have shown that a soft substrate has a significant effect on morphology and cytoskeleton of rat bone marrow mesenchymal stem cel.
OBJECTIVE:To explore the effect of polyacrylamide gels as soft substrates with different elastic moduli on the chondrogenic differentiation of human synovial-derived mesenchymal stem cels.
METHODS:The synovium was harvested from patients with osteoarthritis under sterile conditions, and primary human synovial-derived mesenchymal stem cels were separated using limiting dilution assay. The flow cytometry and multi-directional differentiation experiments were used to identify the cel surface markers and function of the human synovial-derived mesenchymal stem cels, respectively. The polyacrylamide gels with the elastic modulus of 0.4, 6, 30 kPa, which were made using various amounts of acrylamide and bis-acrylamide, were used to culture human synovial-derived mesenchymal stem cels under induction with transforming growth factor-β1 for 7 and 14 days. RT-PCR was used to test the expression of chondrogenic genes, type II colagen gene and cartilage acidic protein 1. The 6-wel cel culture plates served as controls.
RESULTS AND CONCLUSION: The human synovial-derived mesenchymal stem cels showed different cel morphology in the different elastic modulus of polyacrylamide gels. The expression of type II colagen gene and cartilage acidic protein 1 were affected by the different elastic modulus of polyacrylamide gels and culture time, and there was an interaction between these two factors. At 7 days of induction, the expression of cartilage acidic protein 1 gene on 6 kPa polyacrylamide gels was the highest (F=44.350,P=0.000); meanwhile, the expression of type II colagen gene on 0.4 kPa polyacrylamide gels was the highest (F=6.384,P=0.005). These findings indicate that polyacrylamide gels with lower elastic modulus are superior to routine culture plates to promote the chondrogenic differentiation of human synovial-derived mesenchymal stem cels.
