Isolation And Analysis Of Tumor Cells From Human Solid Tumor Tissue Extracted By Needle Biopsy

By David Agorku and Jaak Janssens

Comprehensive molecular analysis of tumor tissue samples is often limited by the amount of tissue available as well as the intratumoral heterogeneity. Needle biopsies yield only small amounts of tissue, which in many cases do not suffice for dissociation and detailed flow cytometric analysis. Additionally, isolation of certain cell populations from biopsy samples for further analysis or cultivation has been extremely challenging. Two major hurdles of needle biopsies are the quality of the cellular material extracted and the potential for lesions within the tissue or the sample.

Among the different biopsy methods and needles available, Spirotome™ Biopsy Needles (Bioncise) have several benefits for sample extraction from various tissues, including high yield^1,2. This is of fundamental importance as a relatively large amount of tissue sample is required for optimal tissue dissociation and subsequent cell isolation, and eventually for representative and reliable results in molecular downstream analyses.

This application note describes a complete workflow ultimately enabling the isolation and analysis of tumor cells from primary breast cancer samples as well as disseminated tumor cells from lymph nodes obtained by biopsy. The workflow includes i) tissue biopsy obtained with Spirotome Needles, ii) dissociation of the solid tumor tissue into single-cell suspensions using the gentleMACS™ Dissociator in combination with the Tumor Dissociation Kit, human and iii) specific isolation of tumor cells using the Tumor Cell Isolation Kit, human. The isolated cells are suitable for different downstream analyses, e.g., flow cytometry, cell culture, or molecular analyses.