CAJ | 학술논문

背景：有研究表明人类软骨糖蛋白39与骨关节软骨的退变与修复具有一定的关系，但其具体的作用机制并不十分明确。目的：观察人类软骨糖蛋白39对成人膝关节软骨祖母细胞成软骨诱导分化的影响。方法：取成人关节软骨，消化分离培养关节软骨祖母细胞；流式细胞仪检测传代细胞中能够表达 CD105、CD166的细胞量并进行分离提纯。将分离的软骨祖母细胞采用单层培养法培养，传代培养至第2代后向分离培养所得的软骨祖母细胞中，分别经过含人类软骨糖蛋白39成软骨培养基及普通成软骨诱导培养基的诱导培养14 d后，通过免疫组织化学染色观察经诱导后细胞中Ⅱ型胶原的表达及通过大体组织学观察评估软骨的形成。结果与结论：关节软骨组织中可以分离出能够表达CD105、CD166的关节软骨祖母细胞，软骨祖母细胞经过诱导分化后逐渐聚集并形成结节，经诱导后Ⅱ型胶原免疫细胞化学着色阳性，且经人类软骨糖蛋白39诱导细胞形成的结节更大，Ⅱ型胶原表达更多。结果表明，成人关节软骨中能够分离培养出具有成软骨分化能力的干细胞系细胞即软骨祖母细胞，且能够被定向诱导分化为软骨细胞，人类软骨糖蛋白39对其分化过程具有一定的促进作用。
배경：유연구표명인류연골당단백39여골관절연골적퇴변여수복구유일정적관계，단기구체적작용궤제병불십분명학。목적：관찰인류연골당단백39대성인슬관절연골조모세포성연골유도분화적영향。방법：취성인관절연골，소화분리배양관절연골조모세포；류식세포의검측전대세포중능구표체 CD105、CD166적세포량병진행분리제순。장분리적연골조모세포채용단층배양법배양，전대배양지제2대후향분리배양소득적연골조모세포중，분별경과함인류연골당단백39성연골배양기급보통성연골유도배양기적유도배양14 d후，통과면역조직화학염색관찰경유도후세포중Ⅱ형효원적표체급통과대체조직학관찰평고연골적형성。결과여결론：관절연골조직중가이분리출능구표체CD105、CD166적관절연골조모세포，연골조모세포경과유도분화후축점취집병형성결절，경유도후Ⅱ형효원면역세포화학착색양성，차경인류연골당단백39유도세포형성적결절경대，Ⅱ형효원표체경다。결과표명，성인관절연골중능구분리배양출구유성연골분화능력적간세포계세포즉연골조모세포，차능구피정향유도분화위연골세포，인류연골당단백39대기분화과정구유일정적촉진작용。
BACKGROUND:Studies have shown that human cartilage glycoprotein-39 has a certain relationship to articular cartilage degeneration and repair, but the mechanism of action is not very clear. OBJECTIVE:To investigate the effect of human cartilage glycoprotein-39 on chondrogenesis of precartilaginous stem cels. METHODS: Precartilaginous stem cels were isolated from the adult articular cartilage. Cels which could express CD105 and CD166 were detected using flow cytometry folowed by isolation and purification. Isolated precartilaginous stem cels werecultured using monolayer method, and then, passage 2 cels were cultured in the medium containing human cartilage glycoprotein-39 and normal chondrogenic medium for 14 days, respectively. Immunohistochemical staining was used to observe expression of type II colagen and gross observation was done for evaluation of cartilage formation. RESULTS AND CONCLUSION:The precartilaginous stem cels isolated from the adult articular cartilage could express CD105 and CD166. After induction, differentiated precartilaginous stem cels gradualy gathered and formed nudes. The induced cels were positive for type II colagen; after induction by human cartilage glycoprotein-39, the nodules became larger and the expression of type II colagen was increased. These findings indicate that precartilaginous stem cels with chondrogenic ability can be isolated from the adult articular cartilage, and can be induced to differentiate into chondrocytes, in which human cartilage glycoprotein-39 plays an important role.