文章摘要
胡 浪,李大鹏,黄永辉.正常及退变髓核细胞诱导骨髓间充质干细胞向髓核方向分化效果比较[J].南京医科大学学报,2014,(5):621~626
正常及退变髓核细胞诱导骨髓间充质干细胞向髓核方向分化效果比较
Effects of normal and degenerative nucleus pulposus cells on differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells
投稿时间:2013-11-27  
DOI:10.7655/NYDXBNS20140517
中文关键词: 椎间盘退变  髓核细胞  骨髓间充质干细胞  共培养  分化
英文关键词: Intervertebral disc degeneration  Nucleus pulposus cells  Bone mesenchymal stem cells  Co-culture  differentiation
基金项目:镇江市科技计划项目(SH2010031);镇江市医学重点人才项目
作者单位
胡 浪 江苏大学临床医学院,江苏 镇江 212013
江苏大学附属医院骨科,江苏 镇江 212001 
李大鹏 江苏大学附属医院骨科,江苏 镇江 212001 
黄永辉 江苏大学附属医院骨科,江苏 镇江 212001 
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中文摘要:
      目的:分离培养SD大鼠退变髓核细胞(degenerative nucleus pulposus cells,dNPCs)?正常髓核细胞(nucleus pulposus cells,NPCs)及骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs),通过Transwell非直接接触体外共培养方法比较dNPCs和NPCs对BMSCs诱导作用?方法:针刺抽吸法制备SD大鼠退变椎间盘动物模型,取正常及已退变的生长良好的第2代NPCs及生长良好的第3代骨BMSCs,将dNPCs 或NPCs等比例与BMSCs分别复合藻酸盐接种于Transwell共培养系统中进行体外非接触共培养(上室接种BMSCs,下室接种dNPCs或NPCs作为dNPCs共培养组和NPCs共培养组)7 d后回收上室BMSCs?单独NPCs和单独BMSCs培养组分别作为阳性和阴性对照?对各组Ⅱ型胶原(collagenⅡ)和多功能蛋白聚糖(aggrecan)行RT-PCR和Western blot检测?结果:collagen Ⅱ和aggrecan的mRNA表达在两共培养组均有较高表达,但NPCs共培养组较dNPCs共培养组更加接近单纯NPCs组,而单纯BMSCs组未见明显表达;collagenⅡ和aggrecan的蛋白表达量在两共培养组中均较高,但NPCs共培养组较dNPCs共培养组更加接近单纯NPCs组,而单纯BMSCs组未见明显表达?结论:在Transwell非直接接触共培养条件下dNPCs和NPCs均能诱导BMSCs向髓核方向分化,并且NPCs对BMSCs向髓核方向分化的诱导作用更加明显?
英文摘要:
      Objective:To isolate and culture the normal and degenerative nucleus pulposus cells (NPCs) and bone mesenchymal stem cells (BMSCs) of SD rats,and then to compare the effects of normal and degenerative NPCs on BMSCs differentiatinto nucleus pulposus-like cells by a 3D noncontact coculture methord. Methods:The animal models with degenerative nucleus pulposus were established by acupuncture and aspiration. The BMSCs,normal and degeneratived NPCs of SD rats were isolated and cultured. The cells growth condition was observed by light microscope,and the second generation of NPCs and dNPCs and the third generation of BMSCs were selecte to combine alginate gel stents and culture in transwell plate. The BMSCs+NPCs or BMSCs+dNPCs were co-cultured in vitro as experiment group;the NPCs and BMSCs were cultured alone as positive and negative control,respectirely. 7 days later,the collagenⅡ and aggrecan of recycled BMSCs of each group were detected by western blot and RT-PCR. Results:The mRNA of collagenⅡand aggrecan showed a high level both in two co-culture groups,but the BMSCs/NPCs co-culture group is more similar to NPCs sample than BMSCs/dNPCs co-culture group. The results of the content of collagenⅡand aggrecan was the same with mRNA level. Conclusion:Under a 3D non-contact co-culture system,both the normal and degenerative NPCs can induce BMSCs differentiation into nucleus pulposus-like cells,but the induction effects of normal NPCs is more obviously than degenerative NPCs .
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