CLIP - Childhood Leukaemia Investigation Prague, Prague, Czech Republic.
Department of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic.
J Neurooncol. 2019 May;143(1):15-25. doi: 10.1007/s11060-019-03135-w. Epub 2019 Mar 2.
The aim of this study was to test the possibility of using specimens obtained by a cavitron ultrasonic surgical aspirator (CUSA) in flow and mass cytometry investigations of pediatric brain tumors.
CUSA specimens obtained from 19 pediatric patients with brain tumors were investigated. Flow and mass cytometry methods were applied to analyze the composition of material collected using the CUSA. Cell suspensions were prepared from CUSA aspirates. Then sample viability was assessed by conventional flow cytometry and subsequently stained with a panel of 31 metal-labeled antibodies.
Viability assessment was performed using conventional flow cytometry. Viability of cells in the acquired samples was below 50% in 16 of 19 cases. A mass cytometry investigation and subsequent analysis enabled us to discriminate brain tumor cells from contaminating leukocytes, whose proportions varied across the specimens. The addition of the viability marker cisplatin directly into the mass cytometry panel gave the means to selecting viable cells only for subsequent analyses. The proportion of non-viable cells was higher among tumor cells compared leukocytes.
When the analysis of the tumor cell immunophenotype is performed with markers for determining viability, the expression of the investigated markers can be evaluated. Suitable markers can be selected by high-throughput methods, such as mass cytometry, and those that are diagnostically relevant can be investigated using flow cytometry, which is more flexible in terms of time.
本研究旨在测试使用 Cavitron 超声外科吸引器（CUSA）获得的标本进行儿科脑肿瘤的流式和液质细胞术研究的可能性。
研究了 19 名脑肿瘤患儿的 CUSA 标本。应用流式和液质细胞术方法分析 CUSA 采集的材料的组成。从 CUSA 吸出物中制备细胞悬浮液。然后，使用传统流式细胞术评估样品的存活率，随后用 31 种金属标记的抗体进行染色。
使用传统流式细胞术进行了存活率评估。在 19 例中有 16 例采集样本中的细胞存活率低于 50%。液质细胞术研究和随后的分析使我们能够区分脑肿瘤细胞与污染的白细胞，其比例在标本之间有所不同。将顺铂等存活标志物直接添加到液质细胞术面板中，可用于仅选择后续分析的活细胞。与白细胞相比，非存活细胞在肿瘤细胞中的比例更高。
当使用确定存活力的标志物对肿瘤细胞免疫表型进行分析时，可以评估所研究标志物的表达。合适的标志物可以通过高通量方法（如液质细胞术）进行选择，并且可以使用更灵活的流式细胞术来研究具有诊断意义的标志物。
