E6流式细胞仪检测CD34^+细胞百分比和绝对计数方法的建立  

Establishment the protocol of CD34^+cell percentage and absolute counting by E6 flow cytometer

作　　者：付笑迎[1] 张小玲[2] 刘梦桐 余阅 陈诗杨 彭冬 邝雅贤 刘四喜[2] 陈运生[1] FU Xiaoying;ZHANG Xiaoling;LIU Mengtong;YU Yue;CHEN Shiyang;PENG Dong;KUANG Yaxian;LIU Sixi;CHEN Yunsheng(Department of Clinical Laboratory,Shenzhen Children's Hospital,Shenzhen,Guangdong,China,518038;Department of Hematology and Oncology,Shenzhen Children's Hospital,Shenzhen,Guangdong,China,518038;Medical College of Jiaying University,Meizhou,Guangdong,China,514031)

机构地区：[1]深圳市儿童医院检验科,广东深圳518038 [2]深圳市儿童医院血液肿瘤科,广东深圳518038 [3]广东嘉应学院医学院,广东梅州514031

出　　处：《分子诊断与治疗杂志》2020年第8期1005-1009,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家自然科学基金(81801588);广东省医学科学技术研究基金项目(A2018329);深圳市三名工程项目(SZSM201512033);深圳小儿血液肿瘤分子医学公共服务平台项目。

摘　　要：目的应用E6流式细胞仪建立检测CD34^+细胞百分比和绝对计数的方法并进行验证。方法收集2019年1月至2019年12月在深圳市儿童医院血液肿瘤科因重型β-地中海贫血行异基因造血干细胞移植的患儿89例,以移植患儿供的骨髓或外周血为研究对象,应用E6流式细胞仪建立ISHAGE法检测CD34^+细胞百分比,应用"流式直接体积法"计算CD34^+细胞的绝对计数,并结合白细胞手工计数对CD34^+细胞绝对计数的结果进行验证。结果结合白细胞手工计数对CD34^+细胞的绝对计数结果的正确性进行验证,Pearson相关分析结果显示:供者外周血干细胞或骨髓干细胞CD34^+细胞绝对计数流式结果与CD34^+细胞绝对计数计算结果高度相关(r=0.93,R2=0.87,P<0.0001);两次供者外周血干细胞或骨髓干细胞白细胞计数和单个核百分比结果高度一致;同样,两次供者外周血干细胞或骨髓干细胞CD34^+细胞流式计数百分比和绝对计数结果亦高度一致。结论应用E6流式细胞仪的"流式直接体积法"计算CD34^+细胞绝对计数结果准确,可以作为造血干细胞移植术前计算供者外周血干细胞或骨髓干细胞CD34^+细胞数目的可靠依据。Objective To establish and verify the method for detecting the percentage and absolute count of CD34^+cells by E6 flow cytometer.Methods A total of 89 children who underwent allogeneic hematopoietic stem cell transplantation for severβ-thalassemia in the Department of Hematology and Oncology of Shenzhen Childrens Hospital from January 2019 to December 2019 were collected,taking bone marrow or peripheral blood from transplanted donor as the research object,the ISHAGE method was established with the E6 flow cytometer for the percentage of CD34^+cells.The absolute count of CD34^+cells was calculated by"flow direct volume method",and validated the result of the absolute count of CD34^+cells with the manual count of white blood cells.Results The correctness of the absolute count of CD34^+cells was validated by combiningthe manual count of white blood cells.The Pearson correlation analysis showed that the absolute count of CD34^+cells in donor peripheral blood stem cells or bone marrow stem cells was highly correlated with the calculated results of absolute CD34^+cell count(r=0.93,R2=0.87,P<0.0001);the white blood cell count and mononuclear percentage results of the peripheral blood or bone marrow stem cells were highly consistent.Similarly,The percentage and absolute count results of CD34^+cells in the peripheral blood stem cells and bone marrow stem cells are also highly consistent.Conclusion The absolute counting results of CD34^+hematopoietic stem cells calculated by the"flow direct volume method"of the E6 flow cytometer are reliable,and can be used as a reliable basis for calculating the number of CD34^+cells in the transplant before hematopoietic stem cell transplantation.

关 键 词：异基因造血干细胞移植 CD34^+细胞 流式细胞术 绝对计数 

分 类 号：R733[医药卫生—肿瘤]