pfc conjugated to texas red  (Celsense Inc)

Journal: Molecular Imaging and Biology

Article Title: Visualizing CAR-T cell Immunotherapy Using 3 Tesla Fluorine-19 MRI

doi: 10.1007/s11307-021-01672-3

Figure Lengend Snippet: Production and characterization of CAR-T cells and their target cells. a Diagrams depicting a lentiviral (LV) tdT-FLuc transfer plasmid co-expressing the tdTomato fluorescent gene and firefly luciferase (FLuc2) bioluminescence reporter gene for the production of NALM6-tdT-FLuc cells and the LV-CD19-CAR-GFP plasmid co-expressing the CD19-targeted second-generation CAR and GFP for the production of CD19 CAR-T cells. b Merged histograms showing the NALM6 cell population before and after transduction with the tdT-FLuc lentivirus. c Bar graph showing the percent expression of CAR/GFP, CD3, CD4, and CD8 markers in CD19-CAR-GFP transduced T cell populations and untransduced T cell populations ( n = 3). d Merged histograms showing a representative CAR-T cell population before and after labeling with red fluorescent perfluorocarbons. d Confocal microscopy shows intracellular localization of red fluorescence in PFC + CAR-T cells and e unlabeled CAR-T cells do not exhibit red fluorescence. These images were merged with brightfield images to show the cell boundary, and blue fluorescence was used to identify CAR-T cell nuclei (DAPI stain).

Article Snippet: To produce PFC-labeled (PFC +) CAR-T cells or PFC + untransduced T cells for 19 F MRI, T cell populations were labeled overnight with 5 mg/ml Texas Red fluorescent dye conjugated PFC (CS-ATM DM Red, Celsense) and washed three times with phosphate-buffered saline (PBS) prior to downstream applications.

Techniques: Plasmid Preparation, Expressing, Luciferase, Transduction, Labeling, Confocal Microscopy, Fluorescence, Staining