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Enrichment of Human Carcinoma Cells for Sensitive Molecular Analysis

Enrichment of Human Carcinoma Cells for Sensitive Molecular Analysis content piece image
Credit: Miltenyi Biotec
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The FFPE Tissue Dissociation Kit by Miltenyi Biotec is designed for the effective dissociation of formalin-fixed paraffin-embedded (FFPE) carcinoma samples for the specific enrichment and subsequent molecular analysis of carcinoma tumor cells. This aims to enhance the sensitivity in molecular experiments, such as mutation analysis by next-generation sequencing (NGS).

Miltenyi Biotec’s FFPE Tissue Dissociation Kit in combination with the gentleMACS™ Octo Dissociator with Heaters provides a standardized protocol for the dissociation of FFPE tissues. In particular, it has been validated for the dissociation of different carcinoma entities for the enrichment of carcinoma tumor cells prior molecular analysis. After dissociation, epitopes of the epithelial cell marker cytokeratin are well preserved. This enables the specific isolation of cytokeratin-positive carcinoma cells from bulk tumor, which can ultimately enhance the sensitivity of molecular analysis of carcinomas.

Benefits of the FFPE Dissociation Kit:

  • Effective enrichment of tumor cells from dissociated carcinoma samples based on cytokeratin expression
  • Enhanced sensitivity of DNA analysis from carcinomas with very low tumor cell content
  • Standardized workflow for reproducible results

FFPE samples are prepared for molecular analysis using a workflow of only four steps: i) deparaffinization and rehydration of FFPE slices; ii) heat-induced antigen retrieval; iii) mechanical and enzymatic dissociation of tumor samples into single-cell suspensions by the gentleMACS™ Octo Dissociator with Heaters and the FFPE Tissue Dissociation Kit, and iv) magnetic enrichment of tumor cells based on cytokeratin expression using the autoMACS® Pro Separator and the Anti-Cytokeratin MicroBeads, human. Both tissue dissociation and magnetic cell enrichment are performed in fully automated processes.

The resulting highly enriched tumor cell fractions are easily amenable to sensitive molecular analysis. The FFPE Dissociation Kit was validated for multiple carcinoma types, including lung, colon, prostate, breast, pancreas, and lymph nodes (metastases).