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Optimization of immunomagnetic selection of myeloma cells from bone marrow using magnetic activated cell sorting

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Abstract

Plasma cells (PCs) enrichment from bone marrow samples of multiple myeloma (MM) patients is frequently performed by immunomagnetic separation (magnetic activated cell sorting, MACS) using anti-CD138 MicroBeads. The aim of our work was to find optimal strategy for immunomagnetic separation of PCs and determine optimal algorithm of separation techniques for samples with various percentage of neoplastic cells. From 2007 to 2008, selection of PCs using separation programs Possels and Posseld2 was carried out on 234 bone marrow samples obtained from 208 MM patients. In 2008, an optimal algorithm for separation programs was introduced based on the analysis of the previous experiments. The Possels program is applicable for samples with >10% PCs in the mononuclear fraction, while the Posseld2 program is used for samples with 5–10% PCs in the mononuclear fraction. Median purity of 92.6% for the positive fraction of cells (range 14.5–99.6%) and median recovery of 60.4% (range 25.7–99.5%) were obtained when the Possels program was applied (n = 45). A total of 80% (36/45) of processed samples had purity of >70%. Median purity for the positive fraction of 83.7% (range 14.3–99.7%) and median recovery of 14.3% (range 3.6–50.0%) were achieved using the Posseld2 program (n = 99). A total of 68% (67/99) of processed samples reached >70% purity. This separation strategy enabled us to obtain sufficient amounts of highly purified PCs required for subsequent research purposes. The MACS method has been unsuccessful if the percentage of PCs in the initial sample was <5%. These samples were processed by fluorescence activated cell sorting (FACS).

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Acknowledgments

We are very grateful to the Department of Internal Medicine and Hematooncology for providing us the bone marrow samples. We thank Dana Kralova for statistical analysis, Jana Moravcova, Marcela Rycova, Drahomira Kyjovska, Bozena Hanakova for MACS separation of samples and Renata Suska, Tomas Perutka for laboratory assistance with FACS analysis. We also thank to all participating members of the Czech Myeloma Group, all patients, their caregivers, and referring physicians for making this work possible. This work was supported by research grants MSMT LC06027, MSM0021622434, IGA NS10408, IGA NS10207, and GACR P304/10/1395.

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Correspondence to Jana Cumova.

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Cumova, J., Kovarova, L., Potacova, A. et al. Optimization of immunomagnetic selection of myeloma cells from bone marrow using magnetic activated cell sorting. Int J Hematol 92, 314–319 (2010). https://doi.org/10.1007/s12185-010-0651-4

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  • DOI: https://doi.org/10.1007/s12185-010-0651-4

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