Establishment of chondrocyte degeneration model in vitro by rat serum
10.3785/j.issn.1008-9292.2015.05.11
- VernacularTitle:应用大鼠血清建立体外软骨细胞退变模型
- Author:
Xiao-Jun WANG
1
;
Hao ZHANG
;
Hong-Sheng ZHAN
;
Dao-Fang DING
Author Information
1. 浙江中医药大学附属湖州市中医院骨科
- Keywords:
Interleukin-1beta;
Chondrocytes;
Rats,wistar;
Serum;
Cell proliferation;
Cartilage diseases;
Cells,cultured
- From:
Journal of Zhejiang University. Medical sciences
2015;(3):308-314
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish a model of chondrocyte degeneration in vitro. Methods:Chondrocytes were isolated from articular cartilages of newly born SD rats by digestion with typeⅡ collagenase .The chondrocytes were cultured with H-DMEM medium containing 10%FBS, 50 ng/mL IL-1β+10%FBS, 2.5% rat serum and 5%rat serum, respectively; and the chondrocytes at passage one were used in the experiments . The morphology changes were investigated under phase contrast microscope after chondrocytes were treated with rat serum and IL-1β.Proliferation of chondrocytes was detected by MTT method .The protein expression levels of PCNA , typeⅡ collagen and MMP-13 were examined by Western blotting . The levels of ADAMTS5, MMP-9, Aggrecan and SOX-9 mRNA were detected by real-time PCR. Results:The cell morphology was changed from polygon to spindle in both rat serum groups and IL-1βgroup, and the proliferation of chondrocytes in these groups was much higher than that in control group .The results showed that the expression levels of typeⅡcollagen, Aggrecan and SOX-9 decreased while the expression levels of MMP-13, MMP-9 and ADMATS5 were up-regulated in rat serum and IL-1β-treated groups compared with control group .Conclusion: The results indicate that rat serum can induce chondrocyte degeneration and may be used for osteoarthritis model in vitro.
