中国组织工程研究与临床康复2011，Vol.15Issue(50)：9323-9326,4.DOI:10.3969/j.issn.1673-8225.2011.50.004
Ⅱ型胶原酶消化法可短时间获得大量纯化大鼠关节软骨细胞
Purified rat chondrocytes can be gained by using typeⅡ collagenase enzyme digestion method in short time
摘要
Abstract
BACKGROUND: To establish an economic, efficient, prjctical isolation and culture system of chondrocytes has importantsignificance to cartilage in witro experiments.OBJECTIVE: To investigate and improve the culture method of rat articutar chondrocytes.METHODS: Rat cartilage of bilateral hip and knee was resected from one week old male rate under aseptic condition.Chondrocytes were isolated by type 0 collagenase enzyme digestion method. The cells were cultured and passaged, thenRESULTS AND CONCLUSION: Inverted phase contrast microscope observation showed that the primary cultured chondrocytes adhcrcntcd at the 12h hour after cultivation. The monolayer formation occurred on the 3rd day after cutivation. And cells were ready to be passaged on the 4h day after cultivation. After passaged for six generations, some cells represented a spindle-Ike appearance. In the 7h generation, most cells turned into a long and irregular appearance, and cell proliferation capacity diminished. Toluidine blue staining showed that the nuclets of cultured chondrocytes were metachromatic Immunofluorescent staining showed that cultured chondrocytes had a positive expression of collagen type 0. These findings illustute that a large number of purified rat chondrocytes can be gained by this method in a short time.关键词
软骨细胞/细胞培养/Ⅱ型胶原酶消化法/鉴定/大鼠分类
医药卫生引用本文复制引用
刘振峰,方锐,孟庆才..Ⅱ型胶原酶消化法可短时间获得大量纯化大鼠关节软骨细胞[J].中国组织工程研究与临床康复,2011,15(50):9323-9326,4.基金项目
新疆维吾尔自治区自治区科技支撑计划项目(编号:200933126),课题名称:维药买朱尼治疗膝骨性关节炎作用机理的研究. （编号:200933126）
