Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads.

IF 5.3 2区医学 Q1 ONCOLOGY Cancer Cell International Pub Date : 2025-01-17 DOI:10.1186/s12935-025-03647-8

Yu Jeong Choi, Jaeguk Choi, Yehyun Kang, Saeam Shin, Seung-Tae Lee, Jong Rak Choi

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Abstract

Background: The prognosis of a plasma cell neoplasm (PCN) varies depending on the presence of genetic abnormalities. However, detecting sensitive genetic mutations poses challenges due to the heterogeneous nature of the cell population in bone marrow aspiration. The established gold standard for cell sorting is fluorescence-activated cell sorting (FACS), which is associated with lengthy processing times, substantial cell quantities, and expensive equipment. Magnetic-activated cell sorting (MACS) can be performed without the need for FACS equipment and allows for rapid sorting of many cells, making it a practical alternative. Our objective is to conduct a comparative analysis of these two sorting techniques to assess whether MACS can viably replace FACS in clinical applications.

Methods: Plasma cell purity, fluorescence in situ hybridization (FISH), and next-generation sequencing analyses were performed on FACS- and MACS-sorted bone marrow samples from 31 PCN patients.

Results: The MACS-sorted samples yielded a higher percentage of plasma cells than FACS-sorted samples under microscopy (p = 0.0156) and flow cytometry (p = 0.0313). FISH performed by two methods in 10 samples showed the same results, and the proportion of abnormal cells was significantly higher in MACS than in FACS (p = 0.001). Wilcoxon matched-pairs signed rank test analysis showed that the median of differences of variant allele frequency (VAF) of two methods (VAF of MACS minus VAF of FACS) in the DNMT3A, TET2, and ASXL1 (DTA) group was - 0.006555 (p = 0.0020), while that in the non-DTA group was 0.002805 (p = 0.0019). Ten copy number variants (CNVs) were found in both FACS- and MACS-sorted samples, eight were identified only in MACS-sorted samples, and one was detected only in FACS-sorted samples.

Conclusion: Our study demonstrates that MACS is a viable alternative for plasma cell sorting in bone marrow samples of patients with PCN.

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评价多发性骨髓瘤患者浆细胞分选方法：流式细胞术与磁珠。

背景：浆细胞肿瘤（PCN）的预后取决于遗传异常的存在。然而，检测敏感的基因突变提出了挑战，由于在骨髓穿刺细胞群的异质性。细胞分选的既定金标准是荧光活化细胞分选（FACS），这与冗长的处理时间、大量的细胞数量和昂贵的设备有关。磁激活细胞分选（MACS）可以在不需要FACS设备的情况下进行，并且允许对许多细胞进行快速分选，使其成为一种实用的替代方案。我们的目的是对这两种分选技术进行比较分析，以评估MACS是否可以在临床应用中替代FACS。方法：对31例PCN患者的FACS和macs分选骨髓标本进行浆细胞纯度、荧光原位杂交（FISH）和下一代测序分析。结果：显微镜下（p = 0.0156）和流式细胞术下（p = 0.0313）， macs分选样品的浆细胞百分比高于facs分选样品。在10个样本中，两种方法的FISH结果相同，MACS中异常细胞的比例明显高于FACS （p = 0.001）。Wilcoxon配对对签名秩检验分析显示，DNMT3A、TET2和ASXL1 （DTA）组两种方法（MACS的VAF减去FACS的VAF）的变异等位基因频率（VAF）差异中位数为- 0.006555 (p = 0.0020)，而非DTA组的差异中位数为0.002805 （p = 0.0019）。在FACS和macs分类的样本中均发现了10个拷贝数变异（CNVs）， 8个拷贝数变异仅在macs分类的样本中被发现，1个拷贝数变异仅在FACS分类的样本中被检测到。结论：我们的研究表明，MACS是PCN患者骨髓样本浆细胞分选的可行选择。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cancer Cell International ONCOLOGY-

CiteScore

10.90

自引率

1.70%

发文量

360

审稿时长

1 months

期刊介绍： Cancer Cell International publishes articles on all aspects of cancer cell biology, originating largely from, but not limited to, work using cell culture techniques. The journal focuses on novel cancer studies reporting data from biological experiments performed on cells grown in vitro, in two- or three-dimensional systems, and/or in vivo (animal experiments). These types of experiments have provided crucial data in many fields, from cell proliferation and transformation, to epithelial-mesenchymal interaction, to apoptosis, and host immune response to tumors. Cancer Cell International also considers articles that focus on novel technologies or novel pathways in molecular analysis and on epidemiological studies that may affect patient care, as well as articles reporting translational cancer research studies where in vitro discoveries are bridged to the clinic. As such, the journal is interested in laboratory and animal studies reporting on novel biomarkers of tumor progression and response to therapy and on their applicability to human cancers.