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Myeloma Bank - Use in Practice



Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Lékařská fakulta

Popis Introduction: Long-term storage and archiving of samples in liquid nitrogen is very important for future use, especially for monitoring and precaution experiments. These samples are also considerable for diagnostic and therapeutic usage in myeloma patients. We present the current status in Myeloma Bank from the year 2001 till presence. Methods/Patients: We collect bone marrow samples from patients with new diagnosis of multiple myeloma (MM), with relapse, before therapy and in the phase of maximal therapeutic response. All patients confirmed the informed consent. We isolate mononuclear cells using gradient centrifugation and then we separate CD138+ (rich in myeloma cells) and CD138- cells with MACS (MAgnetic Cell Sorting, Miltenyi Biotec). We assay purity of these fractions – percentage of myeloma cells on cytospin smear and percentage of CD38+ and CD138+ cells with flow cytometry. Native cell fractions are used in following experiments or they are cryoconserved with programmable freezing and then store in liquid nitrogen. Results: From year 2001 till January 2008 we cryoconserved 8 549 samples from myeloma patients in form of CD138+ and CD138- cell fractions, plasma and bone marrow mononuclear cells. From peripheral blood we store serum, plasma and mononuclear cells. Conclusion: Separated CD138+ cells are used in cytogenetic (FISH, CGH), genomic (RT-PCR) and proteomic experiments. These cells are also exploited as a tumor antigen for vaccine preparation on the dentritic cells basis and for cultivation of myeloma specific T-lymphocytes.
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