Generation and Transduction of NALM6 Cells

NALM6 cells were obtained from ATCC. NALM6 CD19^−/− were generated as previously described^{60 (link)}. NALM6 were transduced to express GFP and firefly Luciferase (FFLuc) for in vitro and in vivo detection^{37 (link)}. Cells were cultured in RPMI 1640 (Corning) with 10% FBS (Hyclone) 1x NEAA, 2mM GlutaMAX, 100U/mL Pen, 100μg/mL Strep, 2mM HEPES (Corning) and 55μM 2-ME). For Incucyte-based analysis, NALM6 CD19⁺ and NALM6 CD19^−/− were transduced with Incucyte NucLight Red lentiviral reagent (NLR, Essen BioScience) and selected with puromycin according to manufacturer’s instructions.

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van der Stegen S.J., Lindenbergh P.L., Petrovic R.M., Xie H., Diop M.P., Alexeeva V., Shi Y., Mansilla-Soto J., Hamieh M., Eyquem J., Cabriolu A., Wang X., Abujarour R., Lee T., Clarke R., Valamehr B., Themeli M., Riviere I, & Sadelain M. (2022). Generation of T-cell-receptor-negative CD8αβ-positive CAR T cells from T cell-derived induced pluripotent stem cells. Nature biomedical engineering, 6(11), 1284-1297.

Publication 2022

RPMI 1640 GlutaMAX HEPES

Cells Firefly luciferase Hepes Puromycin Strep

Corresponding Organization :

Other organizations : Memorial Sloan Kettering Cancer Center, Fate Therapeutics (United States), Amsterdam University Medical Centers, Vrije Universiteit Amsterdam

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Variable analysis

independent variables

Genetic modification: NALM6 CD19-/- cells were generated
Genetic transduction: NALM6 cells were transduced to express GFP and firefly Luciferase

dependent variables

In vitro and in vivo detection of GFP and firefly Luciferase expression

control variables

Cell culture conditions: RPMI 1640 with 10% FBS, 1x NEAA, 2mM GlutaMAX, 100U/mL Pen, 100μg/mL Strep, 2mM HEPES and 55μM 2-ME
For Incucyte-based analysis: NALM6 CD19+ and NALM6 CD19-/- cells were transduced with Incucyte NucLight Red lentiviral reagent and selected with puromycin

positive controls

NALM6 CD19+ cells

negative controls

NALM6 CD19-/- cells

Annotations

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