Journal: Stem cell research

Article Title: Establishment of a human embryonic stem cell line with homozygous TP53 R248W mutant by TALEN mediated gene editing

doi: 10.1016/j.scr.2018.04.013

Figure Lengend Snippet: Reagents details.

Article Snippet: Pluripotency Markers , Mouse anti-TRA-1-85 Alexa Fluor 555-conjugated , 1:600 , R and D Systems Cat# FAB3195A RRID:AB_663789.

Techniques: Immunocytochemistry, Western Blot, Mutagenesis

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: CRISPR/Cas9-mediated CD147 knockout in the THP-1 cell line. A Schematic diagram of CD147 knockout in the THP-1 cell line using nucleofection (Created by BioRender.com/Mahidol University). B Flow cytometric analysis of CD147 expression in the THP-1 cells before nucleofection, after nucleofection (pre-sorting) and post-sorting

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: CRISPR, Knock-Out, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: Generation of the CD147 CAR lentiviral vectors and lentiviral transduction of CD147 CAR into the CD147 KO THP-1 cells. A Schematic representation of the CGW CD147 CAR plasmid vector and lentiviral vector production. LTR, Long terminal repeat; MND, a modified enhancer/promoter of the murine myeloproliferative sarcoma virus promoter; CAR, chimeric antigen receptor; sp, signal peptide; scFv, single chain fragment variable; TM, transmembrane domain; Co-SD, co-stimulatory signaling domain; mEGFP, monomeric enhanced green fluorescent protein (Created by BioRender.com/Mahidol University). B Representative fluorescent microscopic images of the WT THP-1 and CD147 CAR-transduced THP-1 cells on day 2 post-transduction, scale bar = 200 µm. C Flow cytometric analysis of the CD147 CAR-transduced THP-1 cells on day 5 pre- and post-sorting. D Representivative confocal microscopy images demonstrating the binding of the CD147 scFv molecules to the recombinant CD147Rg in the WT THP-1 cells and the CD147 CAR THP-1 cells. (63 × oil immersion objective of the Zeiss LSM 800 confocal microscope, scale bar = 10 µm)

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: Transduction, Plasmid Preparation, Modification, Virus, Confocal Microscopy, Binding Assay, Recombinant, Microscopy

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: Phagocytosis of zymosan bioparticles. A Schematic diagram illustrating the phagocytosis process of zymosan bioparticles by the wild-type (WT), CD147 KO, and CD147 CAR-macrophages (Created by BioRender.com/Mahidol University). B Representative fluorescent microscopic images depicting the WT, CD147 KO, CD147 CAR macrophages co-cultured with the zymosan bioparticles, scale bar = 100 µm. C Representative histograms and D Bar graph showing phagocytic activity against zymosan bioparticles as determined by flow cytometry. Statistical significance was calculated using multiple unpaired t test analyses, ns indicates P > 0.05. Data are results from three independent experiments

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: Cell Culture, Activity Assay, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: Flow cytometric analysis of THP-1 after polarization toward M1 and M2 subtypes. A Representative flow cytometric analysis of the WT, CD147 KO, and CD147 CAR THP-1 cells after activation with PMA (M0, grey), and polarized into M1 (orange) and M2 (green) macrophages. B Mean fluorescence intensity (MFI) of CD80, HLA-DR, CD163, and CD206 of THP-1 cells after polarization. Surface markers include M1 markers (CD80 and HLA-DR), and M2 markers (CD163 and CD206). Statistical significance was calculated using multiple unpaired t test analyses, ns indicates P > 0.05. Data represent the results from three independent experiments

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: Activation Assay, Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: Phagocytosis of cancer cells. A Flow cytometric analysis of CD147 expression in cancer cell lines, including MCF-7, MDA-MB-231, and K562, compared to the isotype control. B Schematic diagram demonstrating the co-culture experiment of the WT, CD147 KO, and CD147 CAR-M against cancer or normal cells (Created by BioRender.com/Mahidol University). C Live-cell time-lapse phagocytosis assay of the WT and CD147 CAR-M against cancer cell lines captured at 1-h intervals for 24 h. The average total red areas per image (µm 2 ) are plotted. D Representative fluorescent images illustrating the phagocytosis of cancer cells by the WT and CD147 CAR-M in each cancer cell line, scale bar = 100 µm. E The comparison of the red area per image (µm 2 ) at 24 h of co-culture. F TNF-⍺ and IL-6 secretion by the WT and CD147 CAR-M after co-culturing with different cancer cell lines. Statistical significance was calculated using multiple unpaired t test analyses, ns indicates P > 0.05, **indicates P < 0.01, ***indicates P < 0.001. Data represent the results from three independent experiments

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: Expressing, Control, Co-Culture Assay, Phagocytosis Assay, Comparison

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

doi: 10.1007/s00262-024-03759-6

Figure Lengend Snippet: Phagocytosis of normal cells. A Flow cytometric analysis of CD147 expression in normal cells, including PBMCs (CD4, CD8, CD56, and CD14 subsets), and MCF10A cell line compared to the isotype control. B Live-cell time-lapse imaging of phagocytosis by the WT macrophages and CD147 CAR-M targeting normal cells, recorded at 1-h intervals over 24 h. The graph displays the average total red areas per image (µm 2 ) for each condition. C The red area per image (µm 2 ) at 24 h of co-culture. D TNF-⍺ and IL-6 secretion by the WT macrophages and CD147 CAR-M after co-culturing with PBMCs and MCF10A cells. Statistical significance was calculated using multiple unpaired t test analysis, ns indicates P > 0.05. Data are results from three independent experiments

Article Snippet: On day 11 post-nucleofection, the nucleofected cells were stained with human anti-CD147 Alexa Flour 488-conjugated antibody (R&D systems, FAB3195G).

Techniques: Expressing, Control, Imaging, Co-Culture Assay