CAJ | 학술논문

背景:自1967年 Manning 等提出的通过胰蛋白酶和细菌胶原酶联合消化法分离软骨细胞以来,软骨细胞的体外分离培养研究较多,但目前尚无统一标准.目的:研究犬关节软骨细胞在体外分离以及培养的条件,寻求体外扩增软骨细胞较简便、可行、高效的实验方法.方法:取3周龄幼犬关节软骨组织,应用胰酶、胶原酶制定8种消化方法获取软骨细胞,对比不同方法所获取细胞数量及细胞成活率,分离细胞进行原代及传代培养,观察各代软骨细胞形态.结果与结论:在体外分离犬关节软骨细胞的各种方法中,单纯应用Ⅱ型胶原酶消化法获得的软骨细胞数最多,细胞成活率最高.软骨细胞可以通过体外培养获得扩增,并且维持良好的细胞形态及表型,但仅限于5代以内.
배경:자1967년 Manning 등제출적통과이단백매화세균효원매연합소화법분리연골세포이래,연골세포적체외분리배양연구교다,단목전상무통일표준.목적:연구견관절연골세포재체외분리이급배양적조건,심구체외확증연골세포교간편、가행、고효적실험방법.방법:취3주령유견관절연골조직,응용이매、효원매제정8충소화방법획취연골세포,대비불동방법소획취세포수량급세포성활솔,분리세포진행원대급전대배양,관찰각대연골세포형태.결과여결론:재체외분리견관절연골세포적각충방법중,단순응용Ⅱ형효원매소화법획득적연골세포수최다,세포성활솔최고.연골세포가이통과체외배양획득확증,병차유지량호적세포형태급표형,단부한우5대이내.
BACKGROUND: Since Manning et al reported to isolate chondrocytes using trypsin and bacterial col agenase digestion in 1967, in vitro isolation and culture of chondrocytes has been widely studied, but there is stil no unified standard. OBJECTIVE: To study the conditions for in vitro culture and isolation of chondrocytes and to explore a simple, feasible, and efficient experimental method to isolate, culture and proliferate canine chondrocytes in vitro. METHODS: The articular cartilage of 3-week-old puppies were isolated and digested with trypsin and type Ⅱcol agenase for harvesting chondrocytes in different conditions. We compared the number of cel s cel survival rate obtained by different methods. Isolated cel s were passaged and morphology of chondrocytes was observed. RESULTS AND CONCLUSION: The number and survival rate of chondrocytes was highest by using simple type Ⅱcol agenase digestion. Chondrocytes can be amplified through in vitro culture and maintain good cel morphology and phenotype, but can only be passaged less than five passages.