与软骨细胞共培养和条件培养液诱导_省略_间充质干细胞向软骨细胞分化的比较_吕晶

中国组织工程研究与临床康复 第 15 卷 第 19 期 2011–05–07 出版 May 7, 2011 Vol.15, No.19 Journal of Clinical Rehabilitative Tissue Engineering Research 与软骨细胞共培养和条件培养液诱导骨髓间充质干细胞向 软骨细胞分化的比较** 吕 晶1，柯 杰1，王 琳2，张冬梅3，刘 冰1，逄键梁1，邓天政1 A comparative study on bone marrow mesenchymal stem cells differentiated into chondrocytes induced by co-culture with chondrocytes and conditioned medium Lü jing1, Ke Jie1, Wang Lin2, Zhang Dong-mei3, Liu Bing1, Pang Jian-liang1, Deng Tian-zheng1 Abstract BACKGROUND: In vitro differentiation of bone marrow mesenchymal stem cells (BMSCs) is dependent on appropriate culture conditions to a great extent. OBJECTIVE: To study the biological characteristics of chondrocytes differentiated from BMSCs co-cultured with chondrocytes versus cultured in conditioned medium. METHODS: Rat BMSCs and chondrocytes were isolated and cultured in neural induction. Cellular non-contact co-culture induced method and conditioned medium induced method were taken to reduce BMSCs differentiating into chondrocytes. And the biological characteristics of the cells were evaluated by MTT, mitotic cycle, GAG secretion, RT-PCR, cell-staining methods, immunocytochemistry. RESULTS AND CONCLUSION: The biological characters of co-culture induced cells were seemed to be better than those of conditioned medium induced cells, which point out that BMSCs co-cultured with chondrocytes are appropriate as seed cells in cartilage tissue engineering. Lü J, Ke J, Wang L, Zhang DM, Liu B, Pang JL, Deng TZ. A comparative study on bone marrow mesenchymal stem cells differentiated into chondrocytes induced by co-culture with chondrocytes and conditioned medium.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2011;15(19): 3421-3426. [doc.xuehai.net doc.xuehai.net] 1 Department of Stomatology, the General Hospital of the Air Force, Beijing 100142, China; 2 Institute of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, China; 3 Department of Stomatology, Out-patient of the Second Retired Cadres, Logistics of Beijing Military Region, Beijing 100071, China Lü Jing, Attending physician, Department of Stomatology, the General Hospital of the Air Force, Beijing 100142, China greatfriday@hotmail. com Correspondence to: Deng Tian-zheng, Doctor, Attending physician, Department of Stomatology, the General Hospital of the Air Force, Beijing 100142, China dengtzh@163.com Supported by: the National Natural Science Foundation of China, No.30900314*, 81000423* Received: 2010-12-06 Accepted: 2011-01-29 摘要 背景：骨髓间充质干细胞体外转化很大程度上依赖于合适的培养条件。 目的：比较与软骨细胞共培养和条件培养液 2 种不同的诱导方案诱导骨髓间充质干细胞向软骨细胞分化的特点。 方法：分离培养大鼠骨髓间充质干细胞和耳软骨细胞，采用骨髓间充质干细胞与软骨细胞共培养及条件培养液诱导成软骨 的方法，诱导骨髓间充质干细胞向软骨细胞分化。以 MTT 法及流式细胞仪检测细胞活性及周期，糖胺多糖、甲苯胺蓝以及 免疫组化染色检测细胞生物学特性，以 RT-PCR 法检测诱导后的软骨细胞Ⅱ型胶原 RNA 表达情况。 结果与结论：采用共培养方式诱导的软骨细胞，其生物学特性与采用条件培养液诱导的软骨细胞相比，前者优于后者，如 分泌糖胺多糖的能力以及基质分泌量均较高。提示共培养方式诱导的软骨细胞更接近正常软骨细胞，更有利于作为组织工 程软骨的种子细胞。 关键词：共培养；软骨细胞；骨髓间充质干细胞；诱导分化；组织工程 doi:10.3969/j.issn.1673-8225.2011.19.001 吕晶，柯杰，王琳，张冬梅，刘冰，逄键梁，邓天政. 与软骨细胞共培养和条件培养液诱导骨髓间充质干细胞向软骨细胞 分化的比较[J].中国组织工程研究与临床康复，2011，15(19):3421-3426. [doc.xuehai.net doc.xuehai.net] 诱导软骨细胞的差异。 0 引言 1 骨 髓 间 充 质 干 细 胞 (bone marrow stem cells， BMSCs)在某些诱导条件下可定向地向软 骨细胞等中胚层来源的组织细胞分化，并适合 自体移植 ，大量的体外诱导实验已经获得成 功，被认为是一种理想的组织工程种子细胞， 为临床上运用 BMSCs 治疗相关疾病提供了有 效依据[2]， 在应用组织工程技术修复软骨损伤方 面显现出良好前景[3-6]，但仍然存在诱导结果不 理想， 导致最终的组织工程产品不理想的问题。 实验基于上述认识，采用不同诱导方案体外诱 导BMSCs向软骨细胞分化，比较不同诱导方案 ISSN 1673-8225 CN 21-1539/R CODEN: ZLKHAH [1] 材料和方法 设计：细胞水平，对比观察实验。 时间及地点：于2010-01/10在解放军空军 总医院航空医学实验室完成。 材料： 2 周龄雄性 SPF 级新西兰大白兔 4 只，体质量0.4 kg，由解放军总医院动物实验 中 心 提 供 ， 动 物 中 心 许 可 证 号 ： SYXK( 军 ) 2002-004。 实验过程中对动物处置符合2006年科学技 术部发布的《关于善待实验动物的指导性意 见》 。 3421

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