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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
引用本文:胡炜,佟敏,杨涛,熊伟,黄异飞. 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[J]. 中国组织工程研究, 2020, 24(19): 2953-2958. DOI: 10.3969/j.issn.2095-4344.2041
作者姓名:胡炜  佟敏  杨涛  熊伟  黄异飞
作者单位: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;华中科技大学附属同济医院,湖北省武汉市 430074
基金项目: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:文题释义:椎间盘微环境:在正常情况下,髓核组织缺乏血液供应,其营养代谢主要依赖渗透。由于代谢产物等影响,椎间盘组织内部的pH值为6.9-7.2,被认为是人体内最恶劣的环境之一,在这种条件下细胞正常代谢能力受到很大抑制,即使是特化的髓核细胞其代谢率也仅为正常的32%。当椎间盘退变后,葡萄糖及氧浓度进一步下降,乳酸堆积,pH值降低和代谢产物进行性增加。髓核间充质干细胞:经过前期实验发现髓核间充质干细胞经自体移植于椎间盘局部,可以分化为纤维环细胞和髓核细胞,或者通过旁分泌细胞因子来促进组织细胞的再生,但髓核间充质干细胞在椎间盘微环境中存活量级并不清楚。椎间盘源性干细胞和髓核间充质干细胞在椎间盘微环境下的生存变化差异性也还是未知。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。ORCID: 0000-0002-3582-5630(胡炜)中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

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  
收稿时间:2019-06-27

Activity of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells in intervertebral disc niche condition
Hu Wei,Tong Min,Yang Tao,Xiong Wei,Huang Yifei. Activity of intervertebral disc stem cells and nucleus pulposus mesenchymal stem cells in intervertebral disc niche condition[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(19): 2953-2958. DOI: 10.3969/j.issn.2095-4344.2041
Authors:Hu Wei  Tong Min  Yang Tao  Xiong Wei  Huang Yifei
Affiliation:Hospitalof Traditional Chinese Medicine Affiliated to Xinjiang Medical University (XinjiangUygur Autonomous Region Hospital of Traditional Chinese Medicine), Urumqi830000, Xinjiang Uygur Autonomous Region, China; Tongji HospitalAffiliated to Huazhong University of Science and Technology, Wuhan 430074,Hubei Province, China
Abstract: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 intervertebral disc stem cells are more active than nucleus pulposus mesenchymal stem cells.
Keywords:intervertebral disc  intervertebral disc stem cells  nucleus pulposus mesenchymal stem cells  CD29  cell proliferation
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