椎间盘微环境下椎间盘源性干细胞和髓核间充质干细胞的活性  

Activity of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells in intervertebral disc niche condition

作　　者：胡炜[1] 佟敏[1] 杨涛[1] 熊伟[2] 黄异飞[1] Hu Wei;Tong Min;Yang Tao;Xiong Wei;Huang Yifei(Hospital of Traditional Chinese Medicine Affiliated to Xinjiang Medical University(Xinjiang Uygur Autonomous Region Hospital of Traditional Chinese Medicine),Urumqi 830000,Xinjiang Uygur Autonomous Region,China;Tongji Hospital Affiliated to Huazhong University of Science and Technology,Wuhan 430074,Hubei Province,China)

机构地区：[1]新疆医科大学附属中医医院(新疆维吾尔自治区中医医院),新疆维吾尔自治区乌鲁木齐市830000 [2]华中科技大学附属同济医院,湖北省武汉市430074

出　　处：《中国组织工程研究》2020年第19期2953-2958,共6页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金项目(201233146-17)，项目负责人：熊伟，黄异飞~~

摘　　要：背景:椎间盘微环境对干细胞生物学行为具有重要作用,拟通过微环境调节作用来实现不依赖种子细胞的椎间盘组织修复。目的:探讨椎间盘微环境下椎间盘源性干细胞和髓核间充质干细胞活性的差异。方法:健康雄性SD大鼠10只,按照解剖区分离椎间盘骨组织,用Ⅱ型胶原酶消化后进行体外培养椎间盘源性干细胞;分离髓核组织,采用酶消化法体外培养髓核间充质干细胞。将获得的椎间盘源性干细胞和髓核间充质干细胞在体外进行正常条件、椎间盘微环境条件下培养扩增,培养第1-6天采用MTT法测定细胞增殖情况,培养第1,3,6天采用流式细胞技术检测CD29阳性表达水平。结果与结论:①在不同环境的培养条件下,椎间盘源性干细胞和髓核间充质干细胞增殖情况呈相反趋势。在正常培养条件下,椎间盘源性干细胞和髓核间充质干细胞呈增殖状态,第4-6天为对数增殖期,两者差异无显著性意义(P>0.05);在椎间盘微环境条件下,髓核间充质干细胞的增殖活性明显低于椎间盘源性干细胞,差异有显著性意义(P<0.05);②在椎间盘微环境条件下培养第3,6天,椎间盘源性干细胞表面CD29阳性抗原表达水平明显高于髓核间充质干细胞,差异有显著性意义(P<0.05);③结果表明,在椎间盘微环境条件下,髓核间充质干细胞和椎间盘源性干细胞活性受到一定程度的抑制,但椎间盘源性干细胞较髓核间充质干细胞保留更多的细胞活性。BACKGROUND:Disc microenvironment plays an important role in the biological behavior of stem cells.It is proposed that the microenvironment can be used to achieve the repair of disc tissue independent of seed cells by niche regulation.OBJECTIVE:To explore the difference in the activity of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells under the intervertebral disc niche.METHODS:Ten healthy male Sprague-Dawley rats were used.The intervertebral disc bone tissue was isolated according to the anatomical area.Intervertebral disc stem cells were digested with type II collagenase and cultured in vitro.Nucleus pulposus was isolated,and nucleus pulposus mesenchymal stem cells were cultured in vitro using enzyme digestion.Intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells were cultured and amplified in normal condition and intervertebral disc niche.The proliferation curves of cultured cells were measured by MTT method at 1-6 days after culture.The expression level of CD29 was detected by flow cytometry at 1,3 and 6 days.RESULTS AND CONCLUSION:Under different culture conditions,the proliferation of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells showed an opposite trend.Under normal culture conditions,intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells were proliferated logarithmically from day 4 to day 6,with no statistical significance(P>0.05).In the intervertebral disc niche condition,the survival rate of nucleus pulposus mesenchymal stem cells was significantly lower than that of intervertebral disc stem cells(P<0.05).In intervertebral disc niche condition,at 3 and 6 days,the surface positive antigen CD29 of intervertebral disc stem cells was significantly higher than that of nucleus pulposus mesenchymal stem cells(P<0.05).These results suggest that in the intervertebral disc niche condition,the activity of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells is inhibited to some extent,but interver

关 键 词：椎间盘 椎间盘干细胞 髓核间充质干细胞 CD29 细胞增殖 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]