Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads
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...
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2025-01-01
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| Series: | Cancer Cell International |
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| Online Access: | https://doi.org/10.1186/s12935-025-03647-8 |
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| author | Yu Jeong Choi Jaeguk Choi Yehyun Kang Saeam Shin Seung-Tae Lee Jong Rak Choi |
| author_facet | Yu Jeong Choi Jaeguk Choi Yehyun Kang Saeam Shin Seung-Tae Lee Jong Rak Choi |
| author_sort | Yu Jeong Choi |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-41e37cb06d644f4baa235b445a7261d0 |
| institution | Kabale University |
| issn | 1475-2867 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj-art-41e37cb06d644f4baa235b445a7261d02025-01-19T12:39:33ZengBMCCancer Cell International1475-28672025-01-012511710.1186/s12935-025-03647-8Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beadsYu Jeong Choi0Jaeguk Choi1Yehyun Kang2Saeam Shin3Seung-Tae Lee4Jong Rak Choi5Department of Laboratory Medicine, Yonsei University College of MedicineDepartment of Laboratory Medicine, Severance HospitalGraduate School of Medical Science, Brain Korea 21 PLUS Project, Yonsei University College of MedicineDepartment of Laboratory Medicine, Yonsei University College of MedicineDepartment of Laboratory Medicine, Yonsei University College of MedicineDepartment of Laboratory Medicine, Yonsei University College of MedicineAbstract 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.https://doi.org/10.1186/s12935-025-03647-8Plasma cell neoplasmPlasma cell myelomaMultiple myelomaCell sortingFluorescence-activated cell sortingFACS |
| spellingShingle | Yu Jeong Choi Jaeguk Choi Yehyun Kang Saeam Shin Seung-Tae Lee Jong Rak Choi Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads Cancer Cell International Plasma cell neoplasm Plasma cell myeloma Multiple myeloma Cell sorting Fluorescence-activated cell sorting FACS |
| title | Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads |
| title_full | Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads |
| title_fullStr | Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads |
| title_full_unstemmed | Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads |
| title_short | Evaluation of plasma cell sorting methods in multiple myeloma patients: flow cytometry versus magnetic beads |
| title_sort | evaluation of plasma cell sorting methods in multiple myeloma patients flow cytometry versus magnetic beads |
| topic | Plasma cell neoplasm Plasma cell myeloma Multiple myeloma Cell sorting Fluorescence-activated cell sorting FACS |
| url | https://doi.org/10.1186/s12935-025-03647-8 |
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