CAJ | 학술논문

背景:诱导分化的骨髓间充质干细胞或髓核细胞移植都存在一定的局限性,抑制自体髓核细胞的退行性变有望成为未来治疗椎间盘退变的有效方法.目的:通过建立骨髓间充质干细胞和髓核细胞分层共培养模型,观察骨髓间充质干细胞对髓核细胞分化的影响.方法:取传至第4代的骨髓间充质干细胞,分为2组:新式分层培养组将髓核细胞接种于去掉挂臂的Transwell小室,膜孔0.4 μm,其与骨髓间充质干细胞比例为1∶1;传统分层培养组同法将髓核细胞接种于带有挂臂的Transwell小室.同时设立单独髓核细胞培养组,各组均培养7 d.免疫组织化学法检测Ⅱ型胶原的表达,~(35)S放射标记渗入放免定量检测蛋白多糖的合成.结果与结论:与单独髓核细胞培养组比较,传统分层培养组、新式分层培养组Ⅱ型胶原阳性细胞数、蛋白多糖合成量均明显增加(P < 0.05,P < 0.01),且新式分层培养组增高幅度大于传统分层培养组(P < 0.05).提示骨髓间充质干细胞与髓核细胞接触共培养后,能显著增加髓核细胞分化增殖和特征性物质的表达,且新式分层培养环境优于传统分层培养.
배경:유도분화적골수간충질간세포혹수핵세포이식도존재일정적국한성,억제자체수핵세포적퇴행성변유망성위미래치료추간반퇴변적유효방법.목적:통과건립골수간충질간세포화수핵세포분층공배양모형,관찰골수간충질간세포대수핵세포분화적영향.방법:취전지제4대적골수간충질간세포,분위2조:신식분층배양조장수핵세포접충우거도괘비적Transwell소실,막공0.4 μm,기여골수간충질간세포비례위1∶1;전통분층배양조동법장수핵세포접충우대유괘비적Transwell소실.동시설립단독수핵세포배양조,각조균배양7 d.면역조직화학법검측Ⅱ형효원적표체,~(35)S방사표기삼입방면정량검측단백다당적합성.결과여결론:여단독수핵세포배양조비교,전통분층배양조、신식분층배양조Ⅱ형효원양성세포수、단백다당합성량균명현증가(P < 0.05,P < 0.01),차신식분층배양조증고폭도대우전통분층배양조(P < 0.05).제시골수간충질간세포여수핵세포접촉공배양후,능현저증가수핵세포분화증식화특정성물질적표체,차신식분층배양배경우우전통분층배양.
BACKGROUND: There are some limitations in induced differentiation of bone-marrow-derived mesenchymal stem cells (hBMSCs) and transplantation of nucleus pulposus cells (NPCs). Inhibition of autologous NPCs degeneration is expected to become an effective way for treatment of disc degeneration in the future. OBJECTIVE: To establish a layered coculture model of hBMSCs and NPCs in vitro, and to observe the effect of hBMSCs on the differentiation of NPCs. METHODS: The fourth passage of hBMSCs was divided into 2 groups. NPCs in the new-style layer culture group were incubated in Transwell cabin without arm, 0.4-μm membrane well; the proportion to BMSCs was 1: 1. NPCs in the traditional culture group were incubated in Transwell cabin with arm. NPC culture group was set. Each group was incubated for 7 days. Collagen Ⅱ was detected by immunohistochemical method. Aggrecan expression was detected by [~(35)S ]-sulfate uptake. RESULTS AND CONCLUSION: Compared with NPC culture group, collagen Ⅱ and aggrecan expression of NPCs were upregulated most evidently in new-style culture and traditional culture groups (P < 0.05, P < 0.01). The increased range of new-style culture group was greater than the traditional culture group (P < 0.05). These results suggested that coculture of BMSCs and NPCs can significantly increase NPC proliferation and specific substance expression. The surrounding of new-style culture surpassed the traditional culture.