CAJ | 학술논문

目的探讨转化生长因子-β1(TGF-β1)诱导兔骨髓间充质干细胞(BMSCs)向髓核样细胞分化的可能性.方法密度梯度法分离培养兔BMSCs,取BMSCs(P3).实验组:含TGF-β1的诱导培养基,对照组:不含TGF-β1诱导培养基,体外培养两周.阿尔新蓝法(Alcian blue)检测培养基内葡糖胺聚糖(GAG)含量.培养第14天,实时定量聚合酶链反应(Real-time PCR)检测髓核样细胞Ⅱ型胶原(CollagenⅡ)及聚集蛋白多糖(Aggrecan) mRNA基因表达.免疫组织化学法测定CollagenⅡ的含量变化.结果 GAG检测结果显示,第7、10、13天实验组GAG含量均显著高于对照组(P＜0.01).实验组CollagenⅡ免疫组织化学染色阳性,表达量较对照组高.Real-time PCR结果证实:实验组CollagenⅡ和Aggrecan mRNA表达水平较对照组显著增高(P＜0.01).结论 TGF-β1能明显增加BMSCs向髓核样细胞分化的诱导生物活性,促进BMSCs向髓核样细胞定向分化.
목적 탐토전화생장인자-β1(TGF-β1)유도토골수간충질간세포(BMSCs)향수핵양세포분화적가능성.방법 밀도제도법분리배양토BMSCs,취BMSCs(P3).실험조:함TGF-β1적유도배양기,대조조:불함TGF-β1유도배양기,체외배양량주.아이신람법(Alcian blue)검측배양기내포당알취당(GAG)함량.배양제14천,실시정량취합매련반응(Real-time PCR)검측수핵양세포Ⅱ형효원(CollagenⅡ)급취집단백다당(Aggrecan) mRNA기인표체.면역조직화학법측정CollagenⅡ적함량변화.결과 GAG검측결과현시,제7、10、13천실험조GAG함량균현저고우대조조(P＜0.01).실험조CollagenⅡ면역조직화학염색양성,표체량교대조조고.Real-time PCR결과증실:실험조CollagenⅡ화Aggrecan mRNA표체수평교대조조현저증고(P＜0.01).결론 TGF-β1능명현증가BMSCs향수핵양세포분화적유도생물활성,촉진BMSCs향수핵양세포정향분화.
Objective To explore the possibility of bone marrow mesenchymal stem cells (BMSCs) stem cells into nucleus pulposus-like cells induced by transforming growth factor-β1 (TGF-β1),thus searching for new treatment for early degenerated intervertebral disc.Methods Density gradient was used for the isolation and culture of rabbit BMSCs,and we collected the third generation BMSCs (P3).BMSCs were cultured for 2 weeks in induction medium containing TGF-β1 (experimental group) and without TGF-β1 (control group).Glycosaminoglycan (GAG) content was detected by Alcian blue.After culturing 14 days,collagen Ⅱ and Aggrecan mRNA gene expression were detected by real-time quantitative polymerase chain reaction (Real-time PCR),and the content changes in collagen Ⅱ were determined by immunohistochemistry.Results GAG content was significantly higher in experimental group at day 7,10 and 13 (P ＜0.01).Immunohistochemical staining showed a positive expression (higher than control group) in experimental group after directional induction.The expression of Collagen Ⅱ and Aggrecan mRNA in experimental group was significantly higher than the control group (P ＜0.01).Conclusion TGF-β1 can obviously increase the biological activity of inducing BMSCs into nucleus pulposus-like cells and promote the directional differentiation from BMSCs to nucleus pulposus-like cells.