Liquid Tumors

Liquid Tumors or non-adherent cells
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Liquid tumors such as leukemia, lymphoma, and myeloma are composed of non-adherent cancer cells. These free-floating liquid tumor cells represent an important target in immuno-oncology but assaying suspension cell cultures can be challenging as cells can be easily washed away or disturbed, hindering further measurements. To create more effective strategies for the treatment of blood cancers, accurate assessments of in vitro target-cell killing are critical.

Track liquid tumor cell proliferation in real time
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Many hematological cancer cell lines can be analyzed using the Maestro platform. Using an antibody-tethering technique to immobilize the target cells you can monitor cell growth in real time. 

 

Daudi cells plated on a CytoView-Z plate
Daudi cells proliferation curve with tethering
Density sweep on different cell types

Liquid tumor cell lines can be assayed in real time. (Left) Daudi cells on a CytoView-Z 96 plate as imaged by the Omni. (Middle) Proliferation of antibody-tethered vs non-tethered Daudi cells as measured by the Maestro Z shows the increase in signal achieved with antibody-tethering. (Right) The tethering technique is broadly applicable as demonstrated in liquid tumor cell lines Daudi, K562, and Raji across a range of cell densities. 

 

Measure dynamic dose-dependent killing of liquid tumor cells
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Real-time assessment can track the magnitude and rate of immune cell-mediated killing. Get a more comprehensive understanding of effector and target cell interactions with kinetic endpoints like kill time 50.

Daudi cell cytolysis over time
Daudi cells with KT50
Daudi cells with EC50

Dose-dependent killing of Daudi cells by CD19-targeted CAR T cells. (Left) Cytolysis of 50K Daudi cells dosed with CD19-targeted CAR T cells at different E:T ratios. (Middle) Kill time 50 (KT50) values for each dose. (Right) A dose-response or immune cell-mediated killing at 96 hours. 

 

Validation of an impedance-based potency assay
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Repeatability and precision is crucial for potency testing. In this collaboration with Miltenyi Biotec, Inc, assay validation of liquid tumor cell killing was performed on the Maestro Z. For the full validation app note, click here.​

Cell Killing (Cytolysis) - Plate 1
Cell Killing (Cytolysis) - Plate 2
Cell Killing (Cytolysis) - Plate 3
Legend

To measure the inter-assay reproducibility, cell killing (cytolysis) was measured in real time across three separate plates. CAR T and nontransduced T cells were added to antibody-tethered lymphocytes (Raji).​

72-hr Cytolysis
KT50
Cytolysis Reproducibility Table

Across three plates, %CV of 72 hr. cytolysis and KT50 met the acceptance criteria of <15%.​

Antibody-mediated tethering of target cells is a reproducible means to quantify CAR T efficacy. The Maestro Z platform meets the criteria for repeatability and precision for potency testing when using a tethered liquid tumor cell line. Read the full validation app note here:​

Miltenyi Biotec, Inc. and Axion BioSystems Collaboration App Note

 

Learn more

 

Application Note: Development of an in vitro cell killing assay for liquid tumor cancer cells
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Assay Steps
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Non-adherent cell lines culture protocol for impedance assays

 

Read our cell culture protocol to learn more about tethered cell assays in the Maestro Z.

Download Protocol

 

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