Microfluidic enrichment, isolation and characterization of disseminated melanoma cells from lymph node samples.

摘要: For the first time in melanoma, novel therapies have recently shown efficacy adjuvant therapy setting, which makes companion diagnostics to guide treatment decisions a desideratum. Early spread of disseminated cancer cells (DCC) sentinel lymph nodes (SLN) is indicative poor prognosis melanoma and early DCCs could therefore provide important information about malignant seed. Here, we present strategy for enrichment from SLN suspensions using microfluidic device (Parsortix™, Angle plc). This approach enables detection isolation viable DCCs, followed by molecular analysis identification genetic changes. By optimizing workflow, established protocol allows high recovery DCC patient-derived node (LN) with harvest rates above 60%. We then assessed integrity transcriptome genome individual, isolated DCCs. In LNs patients, detected expression melanoma-associated transcripts including MLANA (encoding MelanA protein), analyzed BRAF NRAS mutational status confirmed origin comparative genomic hybridization. demonstrate feasibility epitope-independent LN Parsortix™ subsequent characterization single ample application fields use or cellular studies personalized medicine.