Establishment of chondrocyte degeneration model in vitro by rat serum.
- Author:
Xiao-jun WANG
1
,
2
;
Hao ZHANG
1
;
Hong-sheng ZHAN
1
;
Dao-fang DING
1
;
Author Information
- Publication Type:Journal Article
- MeSH: ADAM Proteins; metabolism; ADAMTS5 Protein; Aggrecans; metabolism; Animals; Cartilage, Articular; cytology; Cells, Cultured; Chondrocytes; pathology; Collagen Type II; metabolism; Disease Models, Animal; Interleukin-1beta; pharmacology; Matrix Metalloproteinase 13; metabolism; Matrix Metalloproteinase 9; metabolism; Osteoarthritis; pathology; Proliferating Cell Nuclear Antigen; metabolism; RNA, Messenger; Rats; Rats, Sprague-Dawley; SOX9 Transcription Factor; metabolism; Serum; Up-Regulation
- From: Journal of Zhejiang University. Medical sciences 2015;44(3):308-314
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a model of chondrocyte degeneration in vitro.
METHODSChondrocytes 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.
RESULTSThe cell morphology was changed from polygon to spindle in both rat serum groups and IL-1β group, and the proliferation of chondrocytes in these groups was much higher than that in control group. The results showed that the expression levels of typeⅡ collagen, 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.
CONCLUSIONThe results indicate that rat serum can induce chondrocyte degeneration and may be used for osteoarthritis model in vitro.
