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狗骨髓基质细胞的体外诱导成骨潜能研究
Hits: 1857   Download times: 1143   Received:October 29, 2004    
作者Author单位UnitE-Mail
尚显文 SHANG Xian-wen 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China liuriguang5629519@tom.Com 
刘日光 LIU Ri-guang 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China  
尹培荣 YIN Pei-rong 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China  
杨述华 YANG Shu-hua 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China  
易诚青 YI Cheng-qing 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China  
刘建湘 LIU Jian-xiang 贵阳医学院附属医院骨科 贵州贵阳550004 Department Of Orthopaedics,the Affiliated Hospital of Guiyang Medical College,Guizhou Guiyang,550004,China  
期刊信息:《中国骨伤》2005年18卷,第6期,第343-345页
DOI:doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz
基金项目:国家自然科学基金(编号:30170945);;贵州省科学技术基金(编号:3049)


目的:观察狗骨髓基质细胞的生长特点及诱导条件下的成骨能力。

方法:采用成年狗双侧髂骨取材进行骨髓基质细胞培养,应用四甲基偶氮唑蓝微量酶反应比色法观察细胞增殖;以对硝基苯磷酸盐法及骨钙素含量放免测定法测定碱性磷酸酶活性及骨钙素含量;用VonKossa染色法观察矿化结节;用间接免疫荧光染色法测定Ⅰ型胶原。观察在培养液中添加地塞米松、维生素C、β甘油磷酸钠条件下骨髓基质细胞生长及成骨分化情况。

结果:骨髓基质细胞呈成纤维样表现,增殖力强。诱导条件下的细胞碱性磷酸酶活性及骨钙素含量明显增高,Ⅰ型胶原表达增多,10~12d达到高峰,并出现矿化结节。

结论:狗骨髓基质细胞在本实验条件下,成骨能力肯定,增殖能力强,可作为骨组织工程的种子细胞。
[关键词]:骨髓细胞  骨形成  细胞培养
 
Osteogenic potentiality of cultured dogs marrow stromal cells under condition of bone induction
Abstract:

Objective:To investigate the growth and osteogenic potentiality of cultured dog marrow stromal cells(MSCs).

Methods:Marrow stromal cells were obtained from the dual ilium of adult dogs.The adhesive cells were preserved to passage culture.MSCs were cultured in a conditional medium in subculture including dexamethasone,vitamin C,and β-glycerophosphate.The proliferation was observed by MTT.The ALPase activity and BGP content were detected respectively through PNPP and radioimmunoassay.Von Kossa staining was used for the mineral nodule investigation.The collagen type Ⅰ was observed by indirect immunofluorescent method.

Results:The adhesive MSCs had strong capability of proliferation.The mineral nodules were observed on 12th to 14th days later under osteogenic condition.ALPase activity,BGP and collagen type Ⅰ of MSCs increased significantly,and reached to a peak value on 10th to 12th days.

Conclusion:The MSCs obtained in our experiment have a strong activity of proliferation and osteogenic potentiality.This study reveals that the MSCs can be used as the seed cells in the bone tissue engineering.
 
引用本文,请按以下格式著录参考文献:
中文格式:尚显文,刘日光,尹培荣,杨述华,易诚青,刘建湘.狗骨髓基质细胞的体外诱导成骨潜能研究[J].中国骨伤,2005,18(6):343~345
英文格式:SHANG Xian-wen,LIU Ri-guang,YIN Pei-rong,YANG Shu-hua,YI Cheng-qing,LIU Jian-xiang.Osteogenic potentiality of cultured dogs marrow stromal cells under condition of bone induction[J].zhongguo gu shang / China J Orthop Trauma ,2005,18(6):343~345
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