Culture and identification of chondrocytes from human iliac growth-plate in vitro / 中国矫形外科杂志

ABSTRACT

[Objective] To establish a viable and convenient method for human iliac growth-plate chondrocyte culture in vitro and study the biological characters of the chondrocyte.[Method] The chondrocytes from 10 human iliac growth-plate were cultured in monolayer and passaged to observe the changes of cell morphology.The growth kinetics was detected by using MTT colorimetric.Immunocytochemistry and RT-PCR were performed to investigate the expression of collagenⅡat the protein and mRNA level,respectively.[Result](1)The phenotype of chondrocyte was changed from original polygonal shape to fusiform at 4th passage.(2)MTT test showed the growth curve of the second passage cells presented inverted "S",and cells were found logarithmic growth at 4th~8th day reached platform stage at 9th~10th day,and decreased at the 11th day.(3)The collagen typeⅡimmunohistochemical staining was extensive positive in cells at P2.(4)RT-PCR confirmed that the mRNA level decreased with the cells passaged after P2.[Conclusion]The modified method of the enzymatic two-step digestion is an effective and simple procedure.The second passage cells maintain the phenotype of chondrocyte well,could be serving as seeding cells.