CAJ | 학술논문

目的探讨通过"自组装"(Self-Assembly)培养技术,以生长分化因子-5(GDF-5)诱导人骨髓间充质干细胞(hMSCs),分化形成软骨组织的可能性及效果.方法将第3代hMSCs用含GDF-5的软骨诱导液定向诱导培养.3周后重悬细胞,自组装培养.对自组装组织团块经行大体观察、逆转录-聚合酶链反应(RT-PCR)、免疫组织化学、软骨相关染色检测.结果自组装组织团块有类似于软骨的外观,Ⅱ型胶原mRNA表达明显,组织学显示Ⅱ型胶原和蛋白多糖表达阳性.GDF-5诱导组Ⅱ型胶原免疫组织化学平均吸光度为(0.1678±0.0222),对照组平均吸光度为(0.0908±0.0145),差异有统计学意义(P＜0.05).结论自组装法培养hMSCs能形成具有软骨分子生物学、组织学和生物力学特性的组织团块,而GDF-5能够增强此过程中细胞的软骨表达.
목적 탐토통과"자조장"(Self-Assembly)배양기술,이생장분화인자-5(GDF-5)유도인골수간충질간세포(hMSCs),분화형성연골조직적가능성급효과.방법 장제3대hMSCs용함GDF-5적연골유도액정향유도배양.3주후중현세포,자조장배양.대자조장조직단괴경행대체관찰、역전록-취합매련반응(RT-PCR)、면역조직화학、연골상관염색검측.결과 자조장조직단괴유유사우연골적외관,Ⅱ형효원mRNA표체명현,조직학현시Ⅱ형효원화단백다당표체양성.GDF-5유도조Ⅱ형효원면역조직화학평균흡광도위(0.1678±0.0222),대조조평균흡광도위(0.0908±0.0145),차이유통계학의의(P＜0.05).결론 자조장법배양hMSCs능형성구유연골분자생물학、조직학화생물역학특성적조직단괴,이GDF-5능구증강차과정중세포적연골표체.
Objective To investigate whether cartilage tissue can be developed by self-assembling process with chondrogenically differentiated human mesenchymal stem cells (hMSCs). Methods hMSCs were cultured and differentiated chondrogenically by growth and differentiation factor-5 (GDF-5) in vitro,then self-assembled to form three-dimensional structures. After 4 weeks of self-assembly, the tissue-engineered constructs were observed and detected morphologically, histologically and histochemically. Results The tissue-engineered constructs were hyaline-like in appearance, and had a conspicuous expression of collagen Ⅱ , aggrecan, GAG and proteoglycan. The mean absorbance value of collagen Ⅱ expression in GDF-5 group was (0. 1678 ± 0. 0222), which was higher than that in control group (0.0908 ±0. 0145 )(P ＜0. 05). Conclusion The constructs had the histological, biochemical, and biomechanical properties approaching those of native articular cartilage. Self-assembled cultures appear promising for tissue engineering of cartilage.