组织工程骨-80 ℃低温保存的初步研究
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作者Author单位AddressE-Mail
蓝旭 LAN Xu 解放军兰州军区总医院脊柱外科,甘肃 兰州 730050 Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China lzzyjw@sina.com 
文益民 WEN Yi-min 解放军兰州军区总医院脊柱外科,甘肃 兰州 730050 Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China  
葛宝丰 GE Bao-feng 解放军兰州军区总医院脊柱外科,甘肃 兰州 730050 Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China  
刘雪梅 LIU Xue-mei 解放军兰州军区总医院脊柱外科,甘肃 兰州 730050 Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China  
期刊信息:《中国骨伤》2008年,第21卷,第1期,第49-51页
DOI:doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz
基金项目:全军医学科学技术”十一五“计划课题基金资助项目(编号: 06MA081)
中文摘要:

目的:了解不同冻存方法对组织工程骨生物活性的影响。

方法:以骨髓基质干细胞(marrow stromal cells,MSCs)复合部分脱蛋白骨培养制备组织工程骨,实验分为:A组,组织工程骨用添加冻存保护剂的保存液保存;B组,组织工程骨用不添加冻存保护剂的保存液保存;C组,组织工程骨未行低温保存;D组,单纯MSCs培养。A组和B组的组织工程骨于-80 ℃深低温保存3个月,3个月后复温冻存的组织工程骨。扫描电镜观察MSCs的黏附和分布情况,MTT法检测细胞活力,对硝基苯磷酸法检测碱性磷酸酶(alkaline phosphatase,ALP)活性,流式细胞仪分析细胞周期。

结果:MSCs在材料表面和孔隙内均可黏附和分布,黏附于材料的细胞活力大小依次为 C组>A组>B组 (P<0.01,P<0.05),黏附于材料的细胞ALP活性大小依次为 C组>A组>B组(P < 0.01) .各组细胞周期未见明显变化,未见异倍体细胞。

结论:选择适宜的冻存保护剂对组织工程骨的生物活性有一定的保护作用。
【关键词】组织工程    低温保存
 
Preliminary study on cryopreservation of tissue engineered bone at -80 ℃
ABSTRACT  

Objective: To study the effects of various methods of cryopreservation on the bioactivity of tissue engineered bone.

Methods: MSCs were cocultured with partialy deproteinised bone to produce tissue engineered bone. The experiment was divided into A,B,C and D group. Group A:Tissue engineered bone was stored in preservation solution with cryopreservation medium. Group B:Tissue engineered bone was stored in preservation solution without cryopreservation medium. Group C:Tissue engineered bone was stored without cryopreservation. Group D:MSCs were cultured without cryopreservation. The tissue engineered bone of group A and B had been cryopreserved at -80 ℃ for three months and thawed three months later. The electronic scanning microscope was used to evaluate the adhesion and distribution of MSCs,cell viability was measured by MTT,ALP activity was detected by p-nitrophosphate,cell cycle was analysed by flow cytometry.

Results: MSCs could adhere to the surface of the material and distribute in the hole of material. The cell viability of MSCs adhered to the material was C>A>B group (P<0.01,P<0.05). The ALP activity of MSCs adhered to material was C>A>B group (P<0.01). The cell cycles of different groups did not change significantly;the abnormal cells were not observed.

Conclusion: The choice of proper cryopreservative solution could optimize the bioactivity of tissue engineered bone.
KEY WORDS  Tissue engineering  Bones  Cryopreservation
 
引用本文,请按以下格式著录参考文献:
中文格式:蓝旭,文益民,葛宝丰,刘雪梅.组织工程骨-80 ℃低温保存的初步研究[J].中国骨伤,2008,21(1):49~51
英文格式:LAN Xu,WEN Yi-min,GE Bao-feng,LIU Xue-mei.Preliminary study on cryopreservation of tissue engineered bone at -80 ℃[J].zhongguo gu shang / China J Orthop Trauma ,2008,21(1):49~51
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