Choi Yu Jeong, Choi Jaeguk, Kang Yehyun, Shin Saeam, Lee Seung-Tae, Choi Jong Rak
Department of Laboratory Medicine, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Korea.
Department of Laboratory Medicine, Severance Hospital, Seoul, Korea.
Cancer Cell Int. 2025 Jan 17;25(1):16. doi: 10.1186/s12935-025-03647-8.
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.
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.
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.
Our study demonstrates that MACS is a viable alternative for plasma cell sorting in bone marrow samples of patients with PCN.
浆细胞肿瘤（PCN）的预后因基因异常情况而异。然而，由于骨髓穿刺中细胞群体的异质性，检测敏感的基因突变具有挑战性。既定的细胞分选金标准是荧光激活细胞分选（FACS），其存在处理时间长、所需细胞数量多以及设备昂贵等问题。磁激活细胞分选（MACS）无需FACS设备即可进行，且能快速分选大量细胞，是一种实用的替代方法。我们的目的是对这两种分选技术进行比较分析，以评估MACS在临床应用中是否能切实替代FACS。
对31例PCN患者经FACS和MACS分选的骨髓样本进行浆细胞纯度、荧光原位杂交（FISH）及二代测序分析。
在显微镜下（p = 0.0156）和流式细胞术检测中（p = 0.0313），MACS分选的样本产生的浆细胞百分比高于FACS分选的样本。对10个样本采用两种方法进行FISH检测，结果相同，且MACS检测出的异常细胞比例显著高于FACS（p = 0.001）。Wilcoxon配对符号秩检验分析显示，在DNMT3A、TET2和ASXL1（DTA）组中，两种方法的变异等位基因频率（VAF）差值中位数（MACS的VAF减去FACS的VAF）为 - 0.006555（p = 0.0020），而在非DTA组中为0.002805（p = 0.0019）。在FACS和MACS分选的样本中均发现了10个拷贝数变异（CNV），仅在MACS分选的样本中鉴定出8个，仅在FACS分选的样本中检测到1个。
我们的研究表明，MACS是PCN患者骨髓样本中浆细胞分选的一种可行替代方法。
