Dissociated Tumor Cells

Introduction

The key to unlocking targeted cancer therapies that will improve patient survival lies in the isolation and analysis of live cells in the tumor microenvironment. As primary cells that offer the closest representation of the original cancerous tissue, dissociated tumor cells (DTC) contain tumor, immune, and support cells. Each cell type plays a distinct role in tumorigenesis. Hence, the preservation of each cell type’s population ratio within the microenvironment is vital to the integrity of any downstream application and analysis.

Working with dissociated tumor cell samples is known to be technically challenging, involves rigorous incubation periods, and is associated with low cellular integrity and viability for downstream assay use. Traditionally, the dissociation process is performed using a combination of enzymatic and mechanical separation, which often result in the damage and degradation of cells. Consequently, DTC samples contain a high percentage of debris that interferes with post-processing analysis.

The LeviCell removes debris and enriches for live cells while preserving each unique cell type’s population ratio within the original tumor microenvironment. The aggravating issue of contamination is eliminated by having the entire dissociation process occur in the LeviCell’s closed, touch-free, self-contained environment. Furthermore, the LeviCell’s ability to produce robust, viable cells and abundant yield removes the tedious need for serial subculturing to enrich the tumor cells, thus reducing the overall financial burden and time investment associated with dissociating tumor cells.

Methods

Levitas Bio developed a revolutionary levitation technology platform that uses less than 1 psi of pressure to enable a completely touch-free, label-free, enzyme-free, three-step dissociation process that produces high DTC yield without compromising or biasing cell type populations in the original microenvironment.

Results

The following results illustrate how the LeviCell removes debris as it enriching for live cells while maintaining cell population representation.

The above plots and tables show pre and post LeviCell FACS and Cellometer results indicating increased viability in the live channel following sorting with the LeviCell. The plots and tables on the left were generated from Endometrial DTCs, while the data on the right is from Bladder DTCs. Samples that were or were not sorted with the LeviCell were stained with DAPI and analyzed by flow cytometry and the results are shown in the FACS plots above. Samples were also quantified using a Cellometer results of which are reported in the tables.

The above graphs show pre- and post- LeviCell FACS runs showing that unique populations are maintained within the same ratios. The left indicates results from Endometrial DTCs, and the graph on the right results from Bladder DTCs.

This graph illustrates how the LeviCell removed a considerable amount of debris from Bladder and Endometrial DTC samples while preserving starting viability (Dark Green), LeviCell enriched viability (Light Green).

Conclusion

Viable DTCs are an integral component of therapeutic, diagnostic, and preclinical cancer research. Current methods used for sorting dissociated tumor cells take a certain level of finesse and technical expertise, time, and ample funding to produce sufficient yield and DTC viabilities that enable the analysis of these rare and sensitive cells.

The LeviCell’s 3-step, hands- and label-free platform enables the enrichment of DTCs while maintaining their gene expression and original sample representation ratio for accurate downstream analysis. This unique combination of technological features enables scientists and researchers to focus their effort and expertise on advancing their life-saving scientific knowledge and discoveries.