使用BD FACSCanto II和BD FACSCalibur流式细胞仪，对BD干细胞计数试剂盒进行多中心评估，用于CD34(+)细胞计数。
Multi-site evaluation of the BD Stem Cell Enumeration Kit for CD34(+) cell enumeration on the BD FACSCanto II and BD FACSCalibur flow cytometers.
作者信息
Preti Robert A, Chan Wai Shun, Kurtzberg Joanne, Dornsife Ronna E, Wallace Paul K, Furlage Rosemary, Lin Anna, Omana-Zapata Imelda, Bonig Halvard, Tonn Torsten
机构信息
Progenitor Cell Therapy, NeoStem, Allendale, New Jersey, USA.
Progenitor Cell Therapy, NeoStem, Allendale, New Jersey, USA.
出版信息
Cytotherapy. 2014 Nov;16(11):1558-1574. doi: 10.1016/j.jcyt.2014.03.006. Epub 2014 Jun 11.
BACKGROUND AIMS
Evaluation of the BD Stem Cell Enumeration Kit was conducted at four clinical sites with flow cytometry CD34(+) enumeration to assess agreement between two investigational methods: (i) the BD FACSCanto II and BD FACSCalibur systems and (ii) the predicate method (Beckman Coulter StemKit and StemTrol, Immunotech SAS, Beckman Coulter, Marseille Cedex 9, France).
METHODS
Leftover and delinked specimens (n = 1032) from clinical flow cytometry testing were analyzed on the BD FACSCanto II (n = 918) and BD FACSCalibur (n = 905) in normal and mobilized blood, frozen and thawed bone marrow and leucopheresis and cord blood anticoagulated with citrate phosphate dextrose, anticoagulant citrate dextrose-solution A, heparin and ethylenediaminetetraacetate, alone or in combination. Fresh leucopheresis analysis addressed site equivalency for sample preparation, testing and analysis.
RESULTS
The mean relative bias showed agreement within predefined parameters for the BD FACSCanto II (-2.81 to 4.31 ±7.1) and BD FACSCalibur (-2.69 to 5.2 ±7.9). Results are reported as absolute and relative differences compared with the predicate for viable CD34(+), percentage of CD34(+) in CD45(+) and viable CD45(+) populations (or gates). Bias analyses of the distribution of the predicate low, mid and high bin values were done using BD FACSCanto II optimal gating and BD FACSCalibur manual gating for viable CD34(+), percentage of CD34(+) in CD45(+) and viable CD45(+). Bias results from both investigational methods show agreement. Deming regression analyses showed a linear relationship with R(2) > 0.92 for both investigational methods.
DISCUSSION
In conclusion, the results from both investigational methods demonstrated agreement and equivalence with the predicate method for enumeration of absolute viable CD34(+), percentage of viable CD34(+) in CD45(+) and absolute viable CD45(+) populations.
背景目的
在四个临床地点使用流式细胞术对BD干细胞计数试剂盒进行评估，以评估两种研究方法之间的一致性：（i）BD FACSCanto II和BD FACSCalibur系统；（ii）对照方法（贝克曼库尔特StemKit和StemTrol，免疫技术公司，贝克曼库尔特，法国马赛塞德克斯9区）。
方法
对临床流式细胞术检测剩余的和脱钩的标本（n = 1032）在BD FACSCanto II（n = 918）和BD FACSCalibur（n = 905）上进行分析，样本包括正常血液和动员血液、冷冻和解冻的骨髓、白细胞分离产物以及用枸橼酸盐磷酸盐葡萄糖、抗凝枸橼酸盐葡萄糖溶液A、肝素和乙二胺四乙酸单独或联合抗凝的脐带血。新鲜白细胞分离产物分析涉及样本制备、检测和分析的位点等效性。
结果
BD FACSCanto II（-2.81至4.31±7.1）和BD FACSCalibur（-2.69至5.2±7.9）的平均相对偏差在预定义参数范围内显示出一致性。结果报告为与对照方法相比，活CD34(+)、CD45(+)中CD34(+)百分比以及活CD45(+)群体（或门）的绝对和相对差异。使用BD FACSCanto II最佳门控和BD FACSCalibur手动门控对活CD34(+)、CD45(+)中CD34(+)百分比以及活CD45(+)进行对照方法低、中、高箱值分布的偏差分析。两种研究方法的偏差结果显示出一致性。德明回归分析表明，两种研究方法均呈现线性关系，R(2)>0.92。
讨论
总之，两种研究方法的结果均表明，在绝对活CD34(+)、CD45(+)中活CD34(+)百分比以及绝对活CD45(+)群体的计数方面，与对照方法具有一致性和等效性。
Zotero 插件