Comparison of Chondrocytes Isolated from Healthy People and Osteoarthritis Patients in vitro

Objective:To study the differences of cultivation and biological characteristics of chondrocytes isolated from health people and osteoarthritis patients.Methods: Two-step enzyme digestion was used to isolate chondrocytes.During the cultivation in vitro,the shape and growth changes of the chondrocytes were observed.MTT assay was performed to observe cell proliferation.Toluidine blue staining and immunohistochemical staining were applied to observe the expression of type II collagen.Flow cytometry was performed to detect cell apoptosis.Results: ①Osteoarthritis chondrocytes was similar to fibroblasts in shape,and grew slower than normal chondrocytes.② MTT showed that the OD value of osteoarthritis chondrocytes of the 2nd,4th and 6th generation were lower than that of normal chondrocytes(P0.05).When in the 2nd generation,type II collagen immunohistochemical staining and toluidine blue O staining were weakly positive.The biological characteristics of cartilage cells disappeared in 4th generation.③ The percentage of apoptotic cells of healthy people were significantly lower than that of osteoarthritis patients in the 2nd and 4th generations(P0.05).④ The expressions of type II collagen and proteoglycan in normal chondrocytes were higher than those of osteoarthritis patients.Conclusion:In vitro cultivation of human osteoarthritis chondrocytes are in accordance with cataplasia,thus it can be a good cell model for osteoarthritis study.