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肉苁蓉含药血清诱导骨髓间充质干细胞向成骨细胞分化的实验研究
Hits: 2037   Download times: 1178   Received:October 29, 2009    
作者Author单位UnitE-Mail
曾建春 ZENG Jian-chun 广州中医药大学第一附属医院, 广东 广州 510405 The first Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,Guangdong,China  
樊粤光 FAN Yue-guang 广州中医药大学第一附属医院, 广东 广州 510405 The first Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,Guangdong,China fanyurguang@21cn.com 
刘建仁 LIU Jian-ren 广州中医药大学第一附属医院, 广东 广州 510405 The first Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,Guangdong,China  
曾意荣 ZENG Yi-rong 广州中医药大学第一附属医院, 广东 广州 510405 The first Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,Guangdong,China  
易春智 YI Chun-zhi 广州中医药大学  
阎亮 YAN Liang 广州中医药大学  
期刊信息:《中国骨伤》2010年23卷,第8期,第606-608页
DOI:10.3969/j.issn.1003-0034.2010.08.015
基金项目:国家自然科学基金(编号:30572399)


目的:研究补肾中药肉苁蓉含药血清对骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)定向分化的影响。

方法:全骨髓培养法获得原代骨髓间充质干细胞,经胰酶消化传代培养至第4代,将第4代BMSCs以5×106接种于2块6孔板,每板3组孔,分为空白组、地塞米松组、肉苁蓉组,2 d后换液,加入诱导液。空白组加入含10%胎牛血清(FBS)的 L-DMEM培养基继续培养,地塞米松组在基础培养基中加入诱导剂(含β-甘油磷酸钠10 mmol/L,地塞米松0.1 μmol/L,维生素C 50 mg/L),肉苁蓉组为含10%肉苁蓉含药血清的L-DMEM培养基。第10天取出1块6孔板做ALKP染色,另一6孔板于第20天做6孔板茜素红染色。

结果:第10天肉苁蓉组及地塞米松组ALKP染色阳性,第12天可见白色钙结节,第20天茜素红染色阳性。

结论:BMSCs在肉苁蓉含药血清诱导下可向成骨细胞分化,分别作为组织工程的种子细胞和诱导因子对治疗骨质疏松、骨折不愈合将有良好的前景。
[关键词]:肉苁蓉  骨髓  间质干细胞  成骨细胞  细胞分化
 
Experimental study of directional differentiation of bone mesenchymal stem cells(BMSCs) to osteoblasts guided by serum containing cistanche deserticola
Abstract:

Objective: To study directional differentiation of BMSCs guided by Desertliving Cistanche (Herba Cistanches) which invigorates the kidney.

Methods: Primary BMSCs were obtained by whole bone marrow culture and subcultured to the fourth generation by trypsin digestion,and than inoculated into two six-well plates at 5×106 cells per milliliter,all the plates were divided into three groups as blank group,Dexamethasone(DXM) group and Herba Cistanches group,three wells in each group,medium were changed at day 2. The blank group were changed with L-DMEM containing 10% FBS. The DXM group were changed with medium containing 10 mmol/L β-sodium glycerophosphate,0.1 μmol/L DXM and 50 mg/L vitamin C. The Herba Cistanches group were changed with medium containing 10% blood serum containing Herba Cistanches and L-DMEM. One of the six-well plates was stained by alkaline phosphatase(AKP) at the tenth day,the other one was stained by alizarin Bordeaux at the twentieth day.

Results: At the tenth day DXM group and Herba Cistanches group were ALKP stained positive;from the 12th day,white calcium nodus could be seen at the surface of the wells;which alizarin stained positive by the twentyth day.

Conclusion: The medium containing Herba Cistanches can guide BMSCs to differentiate into osteoblast,which promises a favorable prospect for the treatment of osteoporosis and bone fracture disunion.
KEYWORDS:Cistanche deserticola  Bone marrow  Mesenchymal stem cells  Osteoblasts  Cell differentiation
 
引用本文,请按以下格式著录参考文献:
中文格式:曾建春,樊粤光,刘建仁,曾意荣,易春智,阎亮.肉苁蓉含药血清诱导骨髓间充质干细胞向成骨细胞分化的实验研究[J].中国骨伤,2010,23(8):606~608
英文格式:ZENG Jian-chun,FAN Yue-guang,LIU Jian-ren,ZENG Yi-rong,YI Chun-zhi,YAN Liang.Experimental study of directional differentiation of bone mesenchymal stem cells(BMSCs) to osteoblasts guided by serum containing cistanche deserticola[J].zhongguo gu shang / China J Orthop Trauma ,2010,23(8):606~608
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