eBook: Cell and Gene Therapy - 10

tem is a simple, robust, and highly sensitive means
to measure biological processes in cancer and T
cell ex vivo co-cultures. In summary, CAR-T Target
Luciferase Reporter Cells from ATCC are well-characterized
tools with high reproducibility for studying
CAR-T biofunction and validating new CAR-T
agents for cancer immunotherapy.
Results with Material and Methods
To generate the CAR-T Target Luciferase Reporter
Cells, we selected human tumor cell lines exhibiting
high endogenous expression of clinically relevant
CAR-T target antigens (CD19, CD20, HER2)
on the cell surface. WIL2-S (ATCC® CRL-8885™), Raji
(ATCC® CCL-86™), Daudi (ATCC® CCL-213™), Farage
(ATCC® CRL-2630™), and BT-474 (ATCC® HTB-20™)
were transduced with a lentiviral plasmid expressing
luciferase under the control of an EF1A promoter.
The genetically modified cells were grown
under antibiotic selection, and single cell sorting
was performed to isolate individual clones that
were verified to stably express luciferase (Table 1).
To demonstrate the performance of the CAR-T Target
Luciferase Reporter Cells, we used cancer and
T cell co-culture experiments. Commercially available
CAR-T cells targeting CD19, CD20, and HER2
were employed in this study; empty vector-transduced
T cells from the same donor were paired as
controls. The cytotoxicity of the CAR-T cells against
target tumor cells was measured using a luciferase
assay, a commercially available potency assay, and
Figure 1. CAR-T Target Luciferase Reporter Cells. Schematic showing CAR-T target cells with expression of CD19-positive WIL2-S-Luc2 and Raji-Luc2,
CD20-positive Daudi-Luc2 and Farage-Luc2, and HER2-positive BT-474-Luc2 being surrounded and attacked by CD19-, CD20-, and
HER2-targeting CAR-T cells, respectively. Created with BioRender.com.
Designation
WIL2-S-Luc2
Raji-Luc2
Daudi-Luc2
Farage-Luc2
BT-474-Luc2
ATCC® No.
Table 1. CAR-T Target Luciferase Reporter Cells
Disease
CRL-8885-LUC2™
CCL-86-LUC2™
CCL-213-LUC2™
CRL-2630-LUC2™
HTB-20-LUC2™
B Cell Lymphoma
Burkitt's Lymphoma
Burkitt's Lymphoma
Non-Hodgkin's B Cell Lymphoma
Breast Ductal Carcinoma
Target
CD19
CD19
CD20
CD20
HER2
10

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