Retronectin-Mediated Transduction of NK Cells

24-well flat-bottom non-tissue culture-treated plates (Greiner Bio-One, Austria) were coated with 30 ug/mL Retronectin (Takara, Kusatsu, Japan) and blocked with 2% human serum albumin (Sanquin, Amsterdam, The Netherlands). pLZRS retroviral supernatant was thawed and spun on Retronectin-coated wells at 3000g for 20 minutes at 4°C. Viral supernatant was then removed and 0.5 x 10⁶ NK cells were added to each well in 1 ml of NK-M. After 24 hours, transduced NK cells were transferred to tissue culture-treated culture plates and further expanded in NK-M. This method was previously described by Morton et al. (5 (link), 19 (link)).

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van Hees E.P., Morton L.T., Remst D.F., Wouters A.K., Van den Eynde A., Falkenburg J.H, & Heemskerk M.H. (2024). Self-sufficient primary natural killer cells engineered to express T cell receptors and interleukin-15 exhibit improved effector function and persistence. Frontiers in Immunology, 15, 1368290.

Publication 2024

Retronectin

Corresponding Organization :

Other organizations : Leiden University Medical Center

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Variable analysis

independent variables

Coating the 24-well plates with 30 μg/mL Retronectin
Blocking the Retronectin-coated plates with 2% human serum albumin
Spinning the pLZRS retroviral supernatant on Retronectin-coated wells at 3000g for 20 minutes at 4°C

dependent variables

Transduction efficiency of NK cells

control variables

Using 24-well flat-bottom non-tissue culture-treated plates (Greiner Bio-One, Austria)
Culturing the transduced NK cells in NK-M medium

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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