[Apoptosis of fetus chondrocytes cultured in vitro and expression of caspase-3].
OBJECTIVE
To investigate apoptosis of chondrocytes cultured in vitro and related expression of caspase-3.


METHODS
Apoptosis of chondrocytes were detected by flow cytometry analysis and TUNEL staining. The expression of caspase-3 was determined by RT-PCR and Western blot, and caspase-3 protein activity was determined by ELISA.


RESULTS
Apoptosis was observed in chondrocytes cultured in vitro from passage 1 to passage 4 at various degrees. The percentage of apoptosis of chondrocytes on day 7 was much higher than that on day 3 (15.7% +/- 0.3% vs 8.9% +/- 0.6%, P < 0.01). caspase-3 mRNA and protein expressed in chondrocytes during whole culture process. Along with the culture time extension in vitro, caspase-3 expression and protein activity up-regulated, coincident with apoptosis of chondrocyte. caspase-3 was activated and a fragment of 20 kDa was detected after 7 days of culture.


CONCLUSIONS
caspase-3 is involved in apoptosis of chondrocytes cultured in vitro.
Xiao
Juan
J
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei, P. R. China 430022.
Cao
Yi-lin
YL
Fan
Hua-hua
HH
D017209:Apoptosis; D002358:Cartilage, Articular / Q000166:cytology; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000201:enzymology; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics; D005333:Fetus; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
D004792:Enzyme Precursors; D012333:RNA, Messenger; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases