摘要
背景:骨髓间充质干细胞的分离、鉴定技术在移植治疗中非常关键。目的:应用Cobe Spectra6.1骨髓处理程序分析骨髓间充质干细胞的分离方法。设计、时间及地点:以细胞为对象的观察性实验,于2005-11/2007-11在北京军区总医院完成。材料:骨髓来自进行自体骨髓干细胞移植治疗的缺血性股骨头坏死患者。方法:应用COBE Spectra血液成分单采系统骨髓单个核细胞分离程序,在已建立的运行参数条件下分离骨髓间充质干细胞。主要观察指标:分离前后进行白细胞分类、细胞计数检测、流式细胞分析CD34、CD133和CD271的表达,计算有核细胞回收率、单个核细胞回收率;统计方法分析CD271+和CD133+表达与细胞形态学分类的相关性。结果:①骨髓分离后,有核细胞、单个核细胞明显高于分离前(P均<0.01),回收率分别为26.6%、40.0%。②分离的细胞产品中,CD271+、CD133+和CD34+表达率分别为0.63%、0.51%和1.17%,回收率则依次为72.8%、61.6%和67.6%。③相关分析显示,分离前后CD271+表达率均与类单核样细胞数目相关(P均<0.01);分离后CD34+表达率与类单核样细胞和类淋巴样细胞数目均相关(P均<0.01)。④偏相关分析显示,CD271与类单核样细胞相关(P均<0.01),CD34与类淋巴样细胞相关(P均<0.01)。结论:应用Cobe Spectra6.1骨髓处理程序分离骨髓干细胞用于临床治疗,可以根据治疗需求选择分离类单核样细胞或类淋巴样细胞,有效提高分离效率。
BACKGROUND： Isolation and determination techniques of bone marrow mesenchymal stem cells （BMSCs） are important during transplantation. OBJECTIVE： To analyze the isolation method of BMSCs by Cobe Spectra 6.1 bone marrow treatment sequence. DESIGN, TIME AND SETTING： The cell experiment was performed at the General Hospital of Beijing Military Area Command of Chinese PLA from November 2005 to November 2007. MATERIALS： Bone marrow was collected from ischemic femoral head necrosis patients undergoing autologous bone marrow stem cell transplantation. METHODS： Using COBE Spectra blood constituent system, mononuclear cells were harvested from bone marrow. BMSCs were harvested under an institutional running parameter condition. MAIN OUTCOME MEASURES： Before and after isolation, white blood cells were classified, detected by cell counting. Flow cytometry was employed to analyze the expression of CD34, CD 133 and CD271. Recovery rates of karyocytes and mononuclear cells were calculated. Correlation of CD271^＋ and CD133^＋ expression to cell morphology was statistically analyzed. RESULTS： After bone marrow isolation, the numbers of karyocytes and mononuclear cells were significantly higher than that before isolation （P 〈 0.01）, and the recovery rates were respectively 26.6% and 40.0%. Expression rates of CD271^＋, CD 133^＋ and CD34^＋ were respectively 0.63%, 0.51% and 1.17%, and the recovery rates were respectively 72.8%, 61.6% and 67.6%. Before and after isolation, the expression rate of CD271^＋ was associated with the number of monocytoid cells （P 〈 0.01）. After isolation, the expression rate of CD34^＋ was correlated to the number of monocytoid ceils and lymphoidocytes （P 〈 0.01）. Partial correlation analysis demonstrated that CD271 was associated with monocytoid cells （P 〈 0.01）, and CD34 was related to lymphoidocyte cells （P 〈 0.01）. CONCLUSION： Using Cobe Spectra 6.1 treatment sequence for harvesting bone marrow stem cells in clinic, we can select to collect
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第38期7460-7464,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
首都医学发展基金课题(2005-3043)~~
