Enhancing FISH Analysis Sensitivity with Highly Purified Multiple Myeloma Cells

FISH analysis is crucial in multiple myeloma diagnostics, but challenges arise due to low plasma cell detection rates. By isolating CD138+ malignant plasma cells with the EasySep system, sensitivity can be improved for more accurate genetic aberration detection in cytogenetic analyses, aiding in patient prognosis and treatment decisions.

• FISH analysis
• Multiple Myeloma
• Sensitivity enhancement
• Purified cells
• Cytogenetic analysis

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1. Enhance Sensitivity of FISH Analysis with Highly Purified Multiple Myeloma Cells Meeting or Location - Date

2. Multiple Myeloma - Background Multiple Myeloma (MM) accounts for 10% of all hematological cancers Caused by the proliferation of a malignant plasma cell clones in the bone marrow Although it can be treated using combinational therapy, there is no cure for this disease Photo Courtesy of Ryan A Knudson, Mayo Clinic

3. Multiple Myeloma - Cytogenetic Analysis Oncogenic plasma cell transformation is caused by known genetic aberrations FISH (Fluorescent in situ hybridization) remains the gold standard for the detection of these aberrations The nature of the genetic aberrations found in the sample have a significant impact on the patient prognosis Photo Courtesy of Ryan A Knudson, Mayo Clinic

4. Multiple Myeloma - Diagnostic Challenges Plasma cells can be extremely rare in bone marrow, making their detection difficult Plasma cells culture can also be a challenge due to their low proliferation index Genetic aberrations are therefore undetectable in a portion of cases evaluated using standard techniques False negative diagnosis Photo Courtesy of Ryan A Knudson, Mayo Clinic

5. Multiple Myeloma - Diagnostic Challenges Performing cytogenetic analysis on a sample enriched for malignant plasma cells has been shown to increase assay sensitivity MM cells can be enriched by targeting the marker CD138 (Syndecan-1), which is uniquely expressed on plasma cells Whole Bone Marrow RoboSep Enriched Sample (CD138+) Photo Courtesy of Greg Ahmann, Mayo Clinic

6. The EasySep System EasySep is an immunomagnetic system for fast and easy isolation of cells HIGH PURITY. Achieve purities of up to 98%. COLUMN-FREE. Avoid the hassle and expense of columns by simply pouring off purified cells. GENTLE. Isolate viable cells that are immediately ready for downstream assays including FISH analysis or flow cytometry VERSATILE. Isolate cells directly from virtually any sample source, including whole blood, bone marrow and PBMCs. FLEXIBLE SAMPLE PROCESSING. Process multiple samples over a wide range of sample sizes or automate with RoboSep

7. EasySep Whole Blood / Bone Marrow CD138+ Selection Kit (#18387) Protocol 4) Top up with buffer and place in EasySep magnet 2) Add EasySep Selection Cocktail CD138+ 15 min 10 min 5 min 1) Prepare cells and add RBC EasySep Lysis buffer 5) Pour-off unwanted cells 3) Add EasySep Magnetic Particles Working Volume: 0.5 - 4.5 ml of whole blood or bone marrow

8. EasySep Whole Blood / Bone Marrow CD138+ Selection Kit (#18387) Results Starting with fresh whole blood spiked with a multiple myeloma cell line, U266, the CD138+ cell content of the isolated fraction typically ranges from 89 - 98%. Starting with fresh whole bone marrow, the isolated fraction typically ranges from 77 - 95% (not shown).

9. Fully Automated Cell Isolation with RoboSep -S Up to 4 different samples 4 Big Easy EasySep magnets Typical positive selection 25 to 850 million cells/quadrant 45 minutes Typical negative selection 25 to 425 million cells/quadrant 20 minutes Different protocols executed at same time positive, negative, sequential, custom human, mouse, NHP, rat, other species

10. RoboSep Easy Sample and Protocol Set-up -S Return after run to collect your separated cells. Select protocol. Load sample, EasySep reagents, buffer and tips in carousel. Press Run .

11. RoboSep -S Benefits for Cytogenetics Labs Saves technician time Processes 4 samples in about an hour with only 5 minutes of hands-on time Avoids sample contamination Samples handled independently using disposable tips to eliminate cross-contamination Stay safe All steps are fully automated to minimize contact with samples

12. Questions? www.stemcell.com techsupport@stemcell.com