Immunomagnetic isolation of myeloma cells by autoMACS
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| Year of publication | 2007 |
| Type | Appeared in Conference without Proceedings |
| MU Faculty or unit | |
| Citation | |
| Description | The standard method used for cell separation is Magnetic Antibody Cell Separation, MACS. In our laboratories, we use autoMACS separator (Miltenyi Biotec, Bergisch Gladbach, Germany) designed for positive selection as well as depletion of magnetically labeled cells. The separator operates with separation programs according to cell abundance and the intensity of marker expression. The separated cells are immediately ready for experiments, cell analysis or further subset sorting. Within minutes can be isolated up to 2x108 pure target cells per sample. AutoMACS allows positive selection and depletion of cells even if the antigen used for selection is expressed at low levels. CD138+ myeloma cells can be isolated from whole blood or bone marrow (BM) or from mononuclear cells prepared using gradient centrifugation from BM of the patients with multiple myeloma. The CD138 antigen, also known as syndecan-1, is found on malignant plasma cells in the BM. The CD138+ myeloma cells are isolated via positive selection of human plasma cells using CD138 micro beads. The sample containing labeled cells is loaded onto an autoMACS column which is placed in a magnetic field. The magnetically labeled CD138+ cells are retained within the column. The unlabeled cells run through as negative fraction. Isolated plasma cells can be used for molecular biology studies such as PCR analysis, microarrays, and protein analysis, and for analysis of cell surface markers of intracellular antigens, for example, immunoglobulin light chains. Using possel d program, we obtained 77,6% cells (8,9-112,4%), average (min-max), with purity of positive fraction 83,8% (63,7-98,9%). In this report, we would like to inform about our achieved results in immunomagnetic separation of myeloma cells from patients using several separation programs. |
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