Nalm6-Fluc-Puro/CD19-KO/HuBCMA-Blast is a clonal population of the B cell precursor leukemia cell line Nalm6 that has been engineered to stably express firefly luciferase and human BCMA, while lacking CD19 expression.
The Nalm6-Fluc-Puro/CD19-KO/HuBCMA-Blast cell line was generated by transducing Imanis Nalm6-Fluc-Puro/CD19-KO cells (CL151) with lentiviral vector encoding human BCMA and the blasticidin resistance gene (LV-SFFV-HuBCMA-PGK-Blast (LV073)). Transduced cells were subsequently selected using blasticidin followed by clonal selection using a methylcellulose based semi-solid medium.
This cell line has been tested for mycoplasma and no mycoplasma has been detected. The parental Nalm6 cell line was authenticated by short tandem repeat (STR) profiling.
Low and high density cell photos (200x). 
The indicated number of cells were placed in wells of a 96-well plate. After the addition of 3 mg/mL d-luciferin, bioluminescence was immediately read using a TECAN plate reader.
Nalm6-Fluc-Puro/CD19-KO/HuBCMA-Blast cells were stained with an anti-HuBCMA (left) or anti-HuCD19 (right) antibody (blue) or isotype control antibody (grey) and analyzed by flow cytometry.
Nalm6-Fluc-Puro/CD19-KO/HuBCMA-Blast cells were stained with an anti-HuCD20 (left), anti-HuCD38 (middle), or anti-HuGPRC5D (right) antibody (blue) or isotype control antibody (grey) and analyzed by flow cytometry.
Complete Growth Medium: ATCC-formulated RPMI-1640 supplemented with 10% fetal bovine serum (FBS), and 1X Penicillin/Streptomycin.
The addition of 1 µg/mL puromycin and 3 μg/mL blasticidin to the complete growth medium is recommended if maintaining the cells in culture for more than two weeks.
Grow cells in the indicated medium, subculturing as needed to maintain a density between 3×105 and 3×106 cells/mL. Passage cells by dilution in fresh medium with regular complete media changes.
These cells are suitable for in vitro and in vivo experimentation.
The luciferase transgene facilitates noninvasive bioluminescent imaging of implanted cells in vivo and cell-based assays in vitro.
Nalm6 cells form tumors and metastases at multiple sites (liver, lymph nodes, spleen, bone marrow) upon implantation into immunocompromised mice.
These cells can be amplified to generate additional frozen stocks and cryopreservation of low passage stocks in a designated cryopreservation medium is recommended.
These cells were generated by transduction with a self-inactivating Risk Group 3 (RG3) lentiviral vector. All work should be performed under appropriate biosafety conditions per institutional guidelines.
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