人关节软骨细胞体外培养的传代特点及鉴定

In vitro culture and identification of human articular chondrocytes

目的研究软骨细胞的传代特点，为软骨细胞移植技术的临床应用提供依据。方法用胰酶和Ⅱ型胶原酶对软骨进行双重消化得到软骨细胞，通过免疫组化对软骨细胞进行鉴定。结果软骨细胞贴壁时间为24～36h，传代培养4～5d，细胞形态以圆形为主，根据免疫组化鉴定，这是一种切实可行的培养细胞的有效方法。结论本研究建立了一种简单易行的软骨细胞分离方法，原代及第2代细胞生长情况良好，5代后细胞表型发生变化。

Objective In vitro culture of human chondrocytes is an important means for the study of cartilage diseases,such as osteoarthritis and patellar chondromalacia, and make possible for repairing cartilage defect and malformation by cartilage grafting. This study was carried out to harvest tumorous chondrocytes by a simple and convenient method and reveal their characteristics of passage. Methods Chondrocytes were obtained by double digestion of cartilage using pancreatin and coUagenase Ⅱ and were identified by immuneohistoehemisty. Reaults The time periods of anchorage and passage culture for chondrocytes were 24 - 36 hours and 4 - 5 days , respectively. The cells were mostly round in form and identified by immuneohistochemistry,This method proved to be effective in culturing chondrecytes. Conclusions A simple and easy method for isolating and culturing chondrocytes has been established. The primary and secondary culture cells grew well but the cells after 5 passages showed change in phenoltype.