转壁生物反应器中动态分化组织工程软骨的实验研究
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作者Author单位AddressE-Mail
吕昌伟 LV Chang wei 第四军医大学西京医院全军骨科研究所,陕西西安710032 The Orthopaedics Research Institute of PLA,Xijing Affiliated Hospital of the Fourth Military Medical University lvchangwei00l@mail.china.com 
胡蕴玉 HU Yun yu 第四军医大学西京医院全军骨科研究所,陕西西安710032 The Orthopaedics Research Institute of PLA,Xijing Affiliated Hospital of the Fourth Military Medical University  
白建萍 BAI Jian ping 第四军医大学西京医院全军骨科研究所,陕西西安710032 The Orthopaedics Research Institute of PLA,Xijing Affiliated Hospital of the Fourth Military Medical University  
廉凯 LIAN Kai 第四军医大学西京医院全军骨科研究所,陕西西安710032 The Orthopaedics Research Institute of PLA,Xijing Affiliated Hospital of the Fourth Military Medical University  
期刊信息:《中国骨伤》2004年,第17卷,第3期,第132-134页
DOI:doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz
基金项目:全军医药卫生科研基金资助重大课题(01Z079)
中文摘要:

目的:在转壁生物反应器动态环境中分化骨髓间充质干细胞(bMSC),制备立体组织工程软骨。

方法:穿刺吸取成年兔骨髓,分离扩增bMSC,复合于纤维蛋白凝胶制成圆柱状三维组织(细胞密度20×107/ml),置于转壁生物反应器中进行分化培养,同时设单纯静止培养组。5周后观测大体形态和组织学形态、Ⅰ和Ⅱ型胶原免疫组织化学表达,测量分化组织的细胞活力。

结果:动态培养可以有效保持材料大体形态,甲苯胺兰染色阳性,无明显Ⅰ型胶原表达,大量表达Ⅱ型胶原,人工组织的细胞活力显著高于单纯静止培养方法。

结论:转壁生物反应器培养明显改善组织工程软骨的活力,有利于进行三维材料体外软骨分化。
【关键词】细胞培养  免疫组织化学  生物反应器  软骨
 
In vitro chondrogenesis of tissue engineered biomaterial in rotating wall vessel bioreactor
ABSTRACT  

Objective:To induce the chondrogenesis of bone marrow derived mesenchymal stem cell in rotating wall vessel (RWV) bioreactor and make tissue engineering cartilage in vitro.

Methods:Bone marrow derived mesenchymal stem cells (bMSC) from adult rabbits were separated by gradient density centrifugation and amplified in plates,and were mixed in fibrin sealant gel at final concentration of 2×107/ml and then incubated in RWV bioreactor at 37 ℃,5%CO2.The parallel static culture was performed as control group.Gross appearance,histological section,immunohistochemistry and cell vitality were investigated after 5 weeks.

Results:RWV bioreactor could preserve better gross appearance of compounded material,the sections revealed that Toluidine blue and Ⅱ collagen stain were obviously positive,and the cell vitality of culture in the RWV was higher than the control.

Conclusion:RWV bioreactor can efficiently improve the cell activity of cartilage,and it is a preferable in vitro method for cartilage differentiation of three dimension biomaterial.
KEY WORDS  Cell culture  Immunohistochemistry  Bioreactors  Cartilage
 
引用本文,请按以下格式著录参考文献:
中文格式:吕昌伟,胡蕴玉,白建萍,廉凯.转壁生物反应器中动态分化组织工程软骨的实验研究[J].中国骨伤,2004,17(3):132~134
英文格式:LV Chang wei,HU Yun yu,BAI Jian ping,LIAN Kai.In vitro chondrogenesis of tissue engineered biomaterial in rotating wall vessel bioreactor[J].zhongguo gu shang / China J Orthop Trauma ,2004,17(3):132~134
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