Comparison of imunomagnetic and fluorescence based method of purification of plasma cells from bone marrow
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| Year of publication | 2008 |
| Type | Conference abstract |
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| Description | Introduction: The research of multiple myeloma (MM) is often limited by low infiltration of plasma cells (PCs). We compared two methods of purification - Magnetic Cell Separation (MACS) and Fluorescence Activated Cell Sorting (FACS). Methods: PCs from bone marrows (BM) of 20 newly diagnosed multiple myeloma patients were purified. Each BM was carefully mixed and divided into two aliquots. The mononuclear fraction (MNC) was acquired from one aliquot by Histopaque density gradient centrifugation and CD 138+ PCs were purified by both MACS positive selection (MACS samples) and sorting on FACS Aria (FACS MNC samples). The other aliquot was erythrolysed by the solution with ammonium chlorite and sorted on FACSAria (FACS lysis samples). The percentage of CD138+ cells (purity) was evaluated by morfology. We compared the methods both in all samples and in three groups of them according to the percentage of CD138+ cells assessed by FC immediatelly before purification: the first group with percentage under 5 % (group I), the second 5 -10 % (group II) and the third over 10% (group III). The differences between individual methods and individual groups were statistically evaluated (nonparametric Kruskall-Wallis ANOVA test and Mann-Whitney test). Results: Medians of purities for all samples were 70,9 % (range 8,6-98,8%) in MACS samples, 98,8% (83,3-100%) in FACS MNC samples and 96,4% (50-100%) in FACS lysis samples. The purities were significantly higher both in FACS lysis (p=0,044) and FACS MNC samples (p<0,001) in comparison with MACS samples. There was no significant difference in purities in sorted cells (p= 0,119). In individual groups however, significant differences were seen only in group I (p=0,005 in FACS MNC and p=0,048 in FACS lysis in comparison with MACS). In groups II and III no significant differences in purity were found. Conclusion: FACS and MACS methods of purification of PCs from BM provided comparable purities in samples with the infiltration over 5% of PCs; in samples with lower infiltration the purities were significantly higher in FACS than in MACS. |
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