PMID- 27526448
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20200325
IS  - 1000-1182 (Print)
IS  - 2618-0456 (Electronic)
IS  - 1000-1182 (Linking)
VI  - 34
IP  - 3
DP  - 2016 Jun
TI  - [Effects of in vitro continuous passaging on the phenotype of mouse hyaline 
      chondrocytes and the balance of the extra- cellular matrix].
PG  - 248-54
AB  - OBJECTIVE: This study aimed to investigate the effects of in vitro continuous 
      passaging on the morphological phenotype and differentiation characteristics of 
      mouse hyaline chondrocytes, as well as on the balance of the extracellular matrix 
      (ECM). METHODS: Enzymatic digestion was conducted to isolate mouse hyaline 
      chondrocytes, which expanded over five passages in vitro. Hematoxylin-eosin stain 
      was used to show the changes in chondrocyte morphology. Semi-quantitative 
      polymerase chain reaction was performed to analyze the mRNA changes in the marker 
      genes, routine genes, matrix metalloproteinases (MMPs), and tissue inhibitors of 
      MMPs (TIMPs) in chondrocytes. Zymography was carried out to elucidate changes in 
      gelatinase activities. RESULTS: After continuous expansion in vitro, the 
      morphology of round or polygonal chondrocytes changed to elongated and spindled 
      shape. The expression of marker genes significantly decreased (P < 0.05), and it 
      was almost negatively expressed by P5 chondrocytes. By contrast, the down 
      regulation of routine genes was insignificant. The gene expression levels of MMPs 
      and TIMPs both decreased (P < 0.05), but the change in MMP-1 and TIMP-1 was not 
      statistically significant (P > 0.05). Meanwhile, the ratio of MMPs/TIMPs was 
      altered. At the protein level, the activities of gelatinases decreased after 
      passaging, especially for P4 and P5 chondrocytes (P < 0.05). CONCLUSION: Serially 
      passaged chondrocytes dedifferentiated and lost specific phenotypic 
      characteristics during in vitro expansion culture. Simultaneously, the anabolism 
      and catabolism of the cartilage ECM became uncontrollable and led to the 
      imbalance of ECM homeostasis. When hyaline chondrocytes are applied in research 
      on relevant diseases or cartilage tissue engineering, P0-P2 chondrocytes should 
      be used.
FAU - Linyi, Cai
AU  - Linyi C
FAU - Xiangli, Kong
AU  - Xiangli K
FAU - Jing, Xie
AU  - Jing X
LA  - chi
PT  - Journal Article
PL  - China
TA  - Hua Xi Kou Qiang Yi Xue Za Zhi
JT  - Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal 
      of stomatology
JID - 9422648
RN  - 0 (RNA, Messenger)
RN  - 0 (TIMP1 protein, human)
RN  - 0 (Tissue Inhibitor of Metalloproteinase-1)
RN  - 0 (Tissue Inhibitor of Metalloproteinases)
RN  - EC 3.4.24.- (Gelatinases)
RN  - EC 3.4.24.- (Matrix Metalloproteinases)
RN  - EC 3.4.24.7 (MMP1 protein, human)
RN  - EC 3.4.24.7 (Matrix Metalloproteinase 1)
SB  - IM
MH  - Animals
MH  - Cartilage
MH  - Cell Differentiation
MH  - Cells, Cultured
MH  - Chondrocytes/*physiology
MH  - Cytoskeleton
MH  - Extracellular Matrix
MH  - Gelatinases
MH  - Gene Expression
MH  - Hyalin/*physiology
MH  - Matrix Metalloproteinase 1
MH  - Matrix Metalloproteinases
MH  - Mice
MH  - RNA, Messenger
MH  - Tissue Engineering
MH  - Tissue Inhibitor of Metalloproteinase-1
MH  - Tissue Inhibitor of Metalloproteinases
PMC - PMC7030832
EDAT- 2016/08/17 06:00
MHDA- 2016/12/15 06:00
PMCR- 2016/06/01
CRDT- 2016/08/17 06:00
PHST- 2016/08/17 06:00 [entrez]
PHST- 2016/08/17 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
PHST- 2016/06/01 00:00 [pmc-release]
AID - wcjs-34-03-248 [pii]
AID - 10.7518/hxkq.2016.03.007 [doi]
PST - ppublish
SO  - Hua Xi Kou Qiang Yi Xue Za Zhi. 2016 Jun;34(3):248-54. doi: 
      10.7518/hxkq.2016.03.007.