Biotherapeutics group, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK.
Biosystems and Biomaterials Division, National Institute of Standards and Technology (NIST), Gaithersburg, Maryland, 20899, United States of America.
Cytometry B Clin Cytom. 2019 Nov;96(6):508-513. doi: 10.1002/cyto.b.21773. Epub 2019 Feb 20.
Over 2,000 people a year in the United Kingdom need a bone marrow or blood stem cell transplant. It is important to accurately quantify the hematopoietic stem cells to predict whether the transplant will be successful in replenishing the immune system. However, they are present at low frequency, which complicates accurate quantification. The current gold standard method is single-platform flow cytometry using internal reference counting beads to determine the concentration of CD34 cells. However, volumetric flow cytometers have the ability to measure the acquisition volume, which removes the need for reference beads for calculation of cell concentrations.
In this study, we compared both methods for calculating CD34 cell concentrations in volumetric cytometers, using either the volume reading or the number of reference beads for calculation. In addition, the uncertainty of measurement for each method was estimated.
The results show that both methods have similar uncertainties of measurement. Regression analysis showed low to no statistical difference in CD34 cell concentrations obtained with each method.
Overall, this study suggests that the volumetric method is a valid approach but that the adoption of this technology may be hindered without some form of external calibration of volume readings to increase confidence in the measurement. © 2019 The Authors. Cytometry Part B: Clinical Cytometry published by Wiley Periodicals, Inc. on behalf of International Clinical Cytometry Society.
英国每年有超过 2000 人需要进行骨髓或造血干细胞移植。准确量化造血干细胞对于预测移植是否成功补充免疫系统非常重要。然而，它们的存在频率很低，这使得准确的量化变得复杂。目前的金标准方法是使用内部参考计数珠的单平台流式细胞术来确定 CD34 细胞的浓度。然而，体积流式细胞仪具有测量采集体积的能力，这消除了计算细胞浓度时对参考珠的需求。
在这项研究中，我们比较了使用体积读数或参考珠数量进行计算时，两种方法在体积细胞仪中计算 CD34 细胞浓度的情况。此外，还估计了每种方法的测量不确定度。
结果表明，两种方法的测量不确定度相似。回归分析显示，每种方法获得的 CD34 细胞浓度差异较低，无统计学意义。
总的来说，这项研究表明，体积法是一种有效的方法，但如果没有对体积读数进行某种形式的外部校准，以增加对测量的信心，那么这项技术的采用可能会受到阻碍。2019 年作者。细胞仪部分 B：临床细胞仪由 Wiley 期刊出版，代表国际临床细胞仪学会出版。
