Nalm6-Fluc-Puro/CD19-KO is a clonal population of the B cell precursor leukemia cell line Nalm6 that have been engineered to stably express firefly luciferase while knocking-out CD19 expression.
The Nalm6-Fluc-Puro/CD19-KO cell line was generated by transfecting Nalm6-Fluc-Puro (Imanis #CL151) cells with Cas9-gRNA ribonucleoprotein complexes targeting human CD19. Individual cell clones were generated using a methylcellulose based semi-solid medium and screened for BCMA expression by flow cytometry. Sequencing of the targeted CD19 region confirmed genetic knock-out in the selected cell clone.
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.
Cell photos taken at 200x magnification
The indicated number of cells were placed in wells of a 96-well plate. After the addition of 3 mg/mL d-luciferin, the plate was immediately imaged using a microplate reader. The total relative light units (RLU) was plotted as a function of cell number.
Nalm6-Fluc-Puro/CD19-KO cells were stained with isotype control antibody (grey) or antibody specific to CD19 (green) and analyzed by flow cytometry.
Nalm6-Fluc-Puro/CD19-KO (green) cells were stained with isotype control antibody (grey) or antibody specific to BCMA (A), CD20 (B), GPRC5D (C), or CD38 (D) and analyzed by flow cytometry.
Imanis cell lines Nalm6-Fluc-Puro
, Nalm6-Fluc-Puro/HuCD19-Puro(mid), Nalm6-Fluc-Puro/HuCD19-Puro(low), and Nalm6-Fluc-Puro/CD19-KO were stained with anti-human CD19 antibody or isotype control (grey) and analyzed by flow cytometry. The antibody binding capacity for each cell line is shown.
CD3+ Jurkat cells expressing a CD19-CAR and GFP (Jurkat-CD19-CAR-GFP) or GFP alone (Jurkat-GFP) were co-cultured with either Nalm6-Fluc-Puro (Imanis #CL151) or Nalm6-Fluc-Puro/CD19-KO (Imanis #CL180) target cells. After 48 hours, cells were analyzed by flow cytometry; CD3+ cells were gated for GFP and CD69, a marker of T-cell activation.
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 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.
Don’t see what you need in our catalog? Our scientific team engineers cell lines, viral vectors, and assays and provides the characterization data you need to move forward with confidence.
