Clinical Research Directorate, Frederick National Laboratory for Cancer Research, 9000 Rockville Pike, Building 10, Room B3B51, Bethesda, MD, 20892, USA.
Molecular Imaging Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Sci Rep. 2022 Jun 1;12(1):9125. doi: 10.1038/s41598-022-13144-x.
Labeling immune cells with zirconium-89 (Zr)-oxine has become a viable method to track cells in vivo by PET in various pre-clinical animal models and in clinical applications. Currently, Zr-oxine cell labeling is performed manually, which requires a highly trained specialist and is prone to human error. As the first phase in developing a fully automated radiolabeling system to address this problem, we assess the use of acoustophoresis cell washing to replace the centrifugal cell washing used in the current Zr-oxine cell radiolabeling procedure. To accomplish this, a cell radiolabeling procedure was developed in which two steps requiring a centrifuge to wash cells were replaced using acoustophoresis cell washing methods. The process was tested using murine EL4 lymphoma and T cells. The centrifuge cell labeling procedure was used as a control to compare the acoustophoresis cell washing procedure. The acoustophoresis method produced radiolabeled cells with similar properties to the centrifugal method when comparing labeling efficiency, labeled specific activity, efficacy of removing unbound Zr-oxine from the suspension, cell viability measured using annexin V/propidium iodide staining and activation function. This suggests that acoustophoresis cell washing can be used in the design of an automated benchtop, good manufacture practice-qualified acoustophoresis cell radiolabeling device.
用 89 锝（Zr）-连氮苯乙酸标记免疫细胞已成为通过正电子发射断层扫描（PET）在各种临床前动物模型和临床应用中追踪体内细胞的可行方法。目前，Zr-连氮苯乙酸细胞标记是手动进行的，这需要高度训练有素的专家，并且容易出现人为错误。作为开发全自动放射性标记系统以解决此问题的第一阶段，我们评估了使用声处理细胞洗涤来替代当前 Zr-连氮苯乙酸细胞放射性标记过程中使用的离心细胞洗涤。为此，开发了一种细胞放射性标记程序，其中使用声处理细胞洗涤方法替代了需要离心洗涤细胞的两个步骤。使用鼠 EL4 淋巴瘤和 T 细胞测试了该过程。以离心细胞标记程序作为对照来比较声处理细胞洗涤程序。当比较标记效率、标记比活度、从悬浮液中去除未结合的 Zr-连氮苯乙酸的效果、使用膜联蛋白 V/碘化丙啶染色和激活功能测量的细胞活力时，声处理方法产生的放射性标记细胞与离心方法具有相似的特性。这表明声处理细胞洗涤可用于自动化台式设备的设计，符合良好生产规范（GMP）的声处理细胞放射性标记设备。
