Laminar Wash Sample Preparation for Tumor Infiltrating Lymphocytes (TILs)

Standardize your solid tumor sample preparation with Laminar Wash technology. Achieve cleaner data from dissociated tumor samples.

Preparing a homogenous single cell suspension from solid tumors is a critical step in flow cytometry or other cell analysis experiments. In addition to the dissociation of the tumor, washing of the resulting suspension is a critical step in sample preparation.

Centrifugation-based washing is a step where errors can easily be made, leading to poor data quality from a solid tissue single cell suspension sample. Centrifuging of cells can lead to damage to the cell membrane, especially if the sample contains a relatively small number of cells, scientists are tempted to centrifuge at higher relative centrifugal forces.

Tumor infiltrating lymphocytes are of particular interest to scientists in cell therapy research. The naturally occurring TILs exist in a complex microenvironment containing the extracellular matrix, blood vessels, and stromal and endothelial components in addition to various immune cells. One common problem with TILs preparation occurs during solid tumor dissociation, whereby the TILs are left in a mixture with tissue debris and dead cells in suspension. Consequently, a preparation of autologous TILs often requires further costly and laborious processing such as density gradient centrifugation, immune cell sorting and enrichment, and dead cell and debris removal in combination with multiple centrifugation steps. These unique set of challenges can be ameliorated by the use of the Laminar Wash™ technology.

More Accurate Tumor Infiltrating Lymphocyte (TIL) Population Data

Laminar Wash™ Increases % Live Cells for TILs and Leads To Higher hCD45+ Population
Laminar Wash™ Increases % Live Cells for TILs and LEADS TO HIGHER hCD45+ POPULATION

A549-inoculated humanized mouse (a) unchallenged tumors (b) challenged tumors

Laminar Wash™ Enables Accurate Identification of Tumor Infiltrating Lymphocyte (TIL) Population
LAMINAR WASH™ ENABLES ACCURATE IDENTIFICATION of TUMOR INFILTRATING LYMPHOCYTE (TIL) POPULATION

(a) Centrifuge wash method – cell loss and mechanical stress through sequential pelleting and resuspension
(b) Laminar Wash™ method – less tumor debris, but higher retention of TILs and better resolution of populations

The Laminar Wash™ system allows live cells to settle by gravity followed by a gentle and continuous flow of a fluid across the wells. Contrary to the conventional centrifugation method, the Laminar Wash™ system can effectively remove the floating debris in suspension while keeping the live cells unperturbed in each well. On the Laminar Wash™ plate, the individual cells are washed in a monolayer instead of forming a single pellet, which helps cell surface architecture and epitopes better retained for improved downstream analysis with flow cytometer. Laminar Wash™ results in healthy, viable, and well defined population of TILs, while enhancing the overall quality of data.

Less Debris from Solid Tumor Samples

Laminar Wash™ Leads to Improved Cell Viability of TILs and Less Cytometer Clogging

FACS plots by (a) centrifuge vs. (b) Laminar Wash™ methods.  The dissociated tumor samples from Raji-inoculated Balb/c mice showed higher viability and increased debris removal with Laminar Wash™, leading to less clogging and more consistent reading.

How are you preparing your single cell sequencing samples?

To learn more about the Laminar Wash systems watch this 4-minute video.