Journal: Cancer Cell

Article Title: Single-cell analysis defines a pancreatic fibroblast lineage that supports anti-tumor immunity

doi: 10.1016/j.ccell.2021.06.017

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD24 (150Nd) clone M1/69 , Fluidigm , 3150009B.

Techniques: Blocking Assay, Virus, Recombinant, Plasmid Preparation, Saline, Lysis, Staining, Reverse Transcription, In Vivo, Electroporation, Mass Cytometry, Conjugation Assay, Illumina Sequencing, Library Quantification, In Vitro, Quantitative RT-PCR, Software, Real-time Polymerase Chain Reaction, Cytometry, Microscopy, Imaging, Spectrophotometry

Journal: Stem Cell Research & Therapy

Article Title: Lumican from DESC spheroids enhances odontoblastic differentiation of SCAPs

doi: 10.1186/s13287-025-04849-7

Figure Lengend Snippet: Characterization of SCAPs and DESCs. a Microscopic image of the apical buds from the lower incisors of 5-7-day-old C57BL/6 mice. b Primary mixed cell population. c Purified SCAPs exhibiting spindle-shaped morphology. d Multidirectional differentiation potential of SCAPs. Alcian Blue, Alizarin Red, and Oil Red O staining confirmed glycosaminoglycan deposition, mineralized nodule formation, and lipid droplet accumulation, indicative of chondrogenic, osteogenic, and adipogenic differentiation, respectively. e , f Flow cytometry analysis revealed CD24, CD133, and CD90 positivity, and CD45 negativity in SCAPs ( n = 3). The raw dot plots are presented in Supplementary Fig. S3. g Purified DESCs displaying cobblestone-like morphology. h Immunofluorescence imaging demonstrated positive K14 expression in DESCs (K14, green; DAPI, blue). i , j Flow cytometry analysis revealed CD29 and CD49f positivity, and CD90 negativity in DESCs ( n = 3). The raw dot plots are presented in Supplementary Fig. S3

Article Snippet: For SCAP analysis, cells were labeled with allophycocyanin (APC)-conjugated CD24 antibody (APC-65121, Proteintech, China, 1:300) as well as FITC-conjugated CD133 (11-1331-82, Thermo Fisher Scientific, USA, 1:500), CD90, and CD45 (MCD4501, Thermo Fisher Scientific, USA, 1:500) antibodies.

Techniques: Purification, Staining, Flow Cytometry, Immunofluorescence, Imaging, Expressing

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 1. CD24 is highly expressed in ovarian cancer cell lines and primary ovarian cancer cells, and can be detected by CD24-AF750 NIR staining. (a) Flow cytometry analysis of EOC cell lines OV-90, Caov-3, COV318 and Skov-3 stained with the monoclonal antibody CD24 conjugated to AF750 (n = 3). Each column represents the mean with SD. (b) CD24 expres- sion was evaluated by flow cytometry, and antigen-binding capacity was quantified for OV-90 compared to monoclonal IgG1 antibody (mAb) control. (c) Validation of target-specific binding. White light (colour) image, near infrared (NIR 800) fluorescent image and pseudo-coloured fluorescence intensity merge image of CD24-AF750 stained EOC cell pellets (25 £ 107 tumour cells, scale bar = 1 cm). (d) Immunohistochemistry of four different patient-derived xenograft (PDX) ovarian tumours, stained with anti-CD24. Immunoreactivity score (IRS) 425 § 096 (scale: 1ow = 1 7 = high). Images were captured at 100£ and 200£ magnification (scale bar = 25 mm).

Article Snippet: The monoclonal mouse anti-human CD24 antibody (clone SN3, cat# MCA1379ELX, RRID: AB_321526, Bio-Rad, Oxfordshire, UK) was conjugated to Alexa Fluor 750 NHS ester using the SAIVITM rapid antibody labelling kit and purified by size exclusion chromatography as described by the manufacturer (cat# S30046, Invitrogen, Carlsbad, USA).

Techniques: Staining, Flow Cytometry, Cytometry, Binding Assay, Control, Biomarker Discovery, Immunohistochemistry, Derivative Assay

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 2. Biodistribution and pharmacokinetics of CD24-AF750. (a) Representative example of longitudinal whole-body fluorescence imaging with CD24-AF750 in an OV-90 subcuta- neous and orthotopic xenograft model, performed at 24-h intervals, and bioluminescence imaging (BLI) for confirmation of tumour specificity. (b) Timepoint analysis of the total fluorescence from CD24-AF750 in the subcutaneous tumour, the orthoptic tumour, liver and kidney (n = 3) identified 24 and 48 h as most advantageous. (c) Intraoperative in vivo images of the CD24-AF750 fluorescence in the primary tumour (marked by the dashed line) at 48 and 72 h, showing white light colour image, NIR fluorescence image and pseudo- coloured fluorescence intensity merge image. PT= primary tumour, L= liver. Tumour to background ratios (TBR) were calculated using the mean fluorescence intensity of muscle tis- sue as background (red circle) (d) Comparison of in vivo primary tumour mean fluorescence intensity (measured in arbitrary units; A.U.) and TBR. (e) Representative colour, NIR 800 and pseudo-coloured fluorescent signal merge image of the primary tumour, kidney and liver (all delineated by the dashed lines) at 48 and 72 h, used for (f) ex vivo MFI measure- ments. Statistical analysis (one-way ANOVA with unpaired MannWhitney U test) with p < 005 (*) were regarded as statistically significant. (Scale bars = 1 cm).

Article Snippet: The monoclonal mouse anti-human CD24 antibody (clone SN3, cat# MCA1379ELX, RRID: AB_321526, Bio-Rad, Oxfordshire, UK) was conjugated to Alexa Fluor 750 NHS ester using the SAIVITM rapid antibody labelling kit and purified by size exclusion chromatography as described by the manufacturer (cat# S30046, Invitrogen, Carlsbad, USA).

Techniques: Drug discovery, Imaging, In Vivo, Comparison, Ex Vivo

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 3. Determination of the optimal dose for CD24-targeted intraoperative imaging. (a) Intraoperative in vivo white light (colour) image, near infrared (NIR 800) fluorescent image and pseudo-coloured fluorescence intensity merge image of representative orthotopic OV-90 xenograft models injected with 1, 2, 3 or 4 mg/g of CD24-AF750 48 h before imaging. (PT= primary tumour, marked by white dashed line, and L = liver). (b) Comparison of CD24-AF750 mean fluorescence intensity (MFI) (p = 0.0288 for 1 mg vs 2 mg; p = 0.0112 for 1 mg vs 3 mg) and tumour to background ratio (TBR) (p = 0.0142 for 1 mg vs 3 mg; p = 0.029 for 1 mg vs 4 mg) of the in vivo primary tumour between the four different doses. (c) Ex vivo colour, NIR 800 and pseudo-coloured fluorescent signal merge image of the primary tumour, kidney and liver (all marked by white dashed line) and (d) comparison of ex vivo MFI of the organs between the four different doses. (p = 0.0222 for 1 mg vs 4 mg). Statistical analysis (one-way ANOVA with unpaired MannWhitney U test) with p < 005 (*) were regarded as statistically significant. (Scale bars = 1 cm).

Article Snippet: The monoclonal mouse anti-human CD24 antibody (clone SN3, cat# MCA1379ELX, RRID: AB_321526, Bio-Rad, Oxfordshire, UK) was conjugated to Alexa Fluor 750 NHS ester using the SAIVITM rapid antibody labelling kit and purified by size exclusion chromatography as described by the manufacturer (cat# S30046, Invitrogen, Carlsbad, USA).

Techniques: Imaging, In Vivo, Injection, Comparison, Ex Vivo

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 4. Survival surgery in OV-90luc+ orthoptic xenografts. (a) Intraoperative white light (colour), near infrared (NIR 800) fluorescent and pseudo-coloured fluorescence intensity merge images from CD24-targeted fluorescence image-guided surgery (FIGS) of the primary ovarian tumour and intra-abdominal metastases. Ex vivo bioluminescence imaging was performed on resected lesions to confirm tumour specificity. (Scale bars = 1 cm). (b) Comparison of the number and total weight of resected metastatic lesions between the CD24 FIGS cohort (n = 8) and white light control surgery (n = 8). (c) Comparison of post-operative bioluminescence signal intensity between CD24-FIGS resected mice (n = 6) and white light control mice (n = 6). Longitudinal whole-body bioluminescence imaging was performed weekly for four weeks post-surgery and the bioluminescent signal was normalised to the presurgical bioluminescence signal of each individual mouse. (d) Mice were allocated a disease stage based on pre-surgical bioluminescence signal and intraoperative disease dissemination (e) Comparison of (normalised) pre- and post-surgical bioluminescence signal between CD24-FIGS and white light surgery in Stage I (f) Stage II and (g) Stage III resected mice (p > 005). Statistical analysis (MannWhitney U test) with p < 005 (*) were regarded as statistically significant.

Article Snippet: The monoclonal mouse anti-human CD24 antibody (clone SN3, cat# MCA1379ELX, RRID: AB_321526, Bio-Rad, Oxfordshire, UK) was conjugated to Alexa Fluor 750 NHS ester using the SAIVITM rapid antibody labelling kit and purified by size exclusion chromatography as described by the manufacturer (cat# S30046, Invitrogen, Carlsbad, USA).

Techniques: Ex Vivo, Imaging, Comparison, Control

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 5. Intraoperative CD24-targeted fluorescence imaging of four heterogeneous PDX of advanced stage HGSOC. (a) Intraoperative white light (colour), near infrared (NIR 800) fluo- rescent and pseudo-coloured fluorescence intensity merge images show positive identification of primary tumour and small metastatic lesions in four separate, orthotopically implanted PDX models of HGSOC, after injection with 3 mg/g CD24-AF750 48 h prior to intraoperative fluorescence imaging. Ex vivo optical imaging confirmed fluorescence specific- ity. Positive lesions are represented by dashed lines in the colour view image. The arrow points to the strongest fluorescent signal in the near infrared (NIR) 800 images. (Scale bar, 1 cm). (b) CD24 expression was confirmed by IHC in fluorescence positive PDX lesions. CD24 staining was evaluated in all five resected lesions with an immunoreactivity score (44 § 11, scale: low 1 7 high). CK8 staining was used to confirm human tumour origin. Magnification 200£. (c) CD24 and folate receptor alpha IHC immunoreactivity score com- parison on PDX tissue sections, demonstrating no significance (p > 005, MannWhitney U test)) difference (n = 21). Images were captured at 100£ and 200£ magnification (scale bar = 25 mm).

Article Snippet: The monoclonal mouse anti-human CD24 antibody (clone SN3, cat# MCA1379ELX, RRID: AB_321526, Bio-Rad, Oxfordshire, UK) was conjugated to Alexa Fluor 750 NHS ester using the SAIVITM rapid antibody labelling kit and purified by size exclusion chromatography as described by the manufacturer (cat# S30046, Invitrogen, Carlsbad, USA).

Techniques: Imaging, Injection, Ex Vivo, Optical Imaging, Expressing, Staining

Journal: STAR Protocols

Article Title: Simultaneous isolation of CD45 tumor-infiltrating lymphocytes, tumor cells, and associated fibroblasts from murine breast tumor model by MACS

doi: 10.1016/j.xpro.2022.101951

Figure Lengend Snippet: Panel for flow cytometry analysis of sorted populations

Article Snippet: CD24 Antibody, anti-mouse, REAfinity-PE-vio770 (1:100 dilution) , Miltenyi Biotec , Cat# 130-110-828, RRID: AB_2656546.

Techniques: Flow Cytometry, Marker, Staining

Journal: STAR Protocols

Article Title: Simultaneous isolation of CD45 tumor-infiltrating lymphocytes, tumor cells, and associated fibroblasts from murine breast tumor model by MACS

doi: 10.1016/j.xpro.2022.101951

Figure Lengend Snippet: Sample detail for fluorescence compensation settings

Article Snippet: CD24 Antibody, anti-mouse, REAfinity-PE-vio770 (1:100 dilution) , Miltenyi Biotec , Cat# 130-110-828, RRID: AB_2656546.

Techniques: Fluorescence, Staining, Suspension

Journal: STAR Protocols

Article Title: Simultaneous isolation of CD45 tumor-infiltrating lymphocytes, tumor cells, and associated fibroblasts from murine breast tumor model by MACS

doi: 10.1016/j.xpro.2022.101951

Figure Lengend Snippet:

Article Snippet: CD24 Antibody, anti-mouse, REAfinity-PE-vio770 (1:100 dilution) , Miltenyi Biotec , Cat# 130-110-828, RRID: AB_2656546.

Techniques: Recombinant, Flow Cytometry, Staining, Isolation, Cell Isolation, Software, Real-time Polymerase Chain Reaction, Blocking Assay, Gentle

Journal: Cancer Cell International

Article Title: Sporadic PCDH18 somatic mutations in EpCAM-positive hepatocellular carcinoma

doi: 10.1186/s12935-017-0467-x

Figure Lengend Snippet: Hepatic stem cell marker expression in HCC1 and HCC2 cells. a Flow cytometry of HCC1 and HCC2 cells using fluorescently-labeled antibodies against EpCAM, CD133, CD90, CD44, CD24, and CD13. b Flow cytometry of EpCAM + cells using an anti-EpCAM antibody. Figure shows EpCAM + HCC1 cells on days 1, 3, and 7 after cell sorting and EpCAM + HCC2 cells on days 1, 7, and 14 after cell sorting. c Histological analysis of EpCAM + HCC1 and HCC2 xenografts. The figure shows hematoxylin and eosin (H&E) staining and anti-EpCAM immunohistochemistry (IHC) staining of the tumors. d Representative phase-contrast images of sorted EpCAM + and EpCAM − HCC1 and HCC2 cell spheroids. e EpCAM + and EpCAM − HCC1 and HCC2 spheroid formation. Experiments were performed in triplicate. Bars indicate the mean ± standard deviation. f Tumorigenic potential of EpCAM + cells. Representative photomicrographs of NOD/SCID mice (upper panel) and subcutaneous tumors (lower panel) from EpCAM + and EpCAM − HCC1 and HCC2 cell xenografts

Article Snippet: The antibodies used were: a FITC-conjugated anti-EpCAM monoclonal antibody (Clone Ber-EP4; DAKO, Carpinteria, CA); an APC-conjugated anti-CD326 (EpCAM) antibody (Miltenyi Biotec K.K., Tokyo, Japan); an APC-conjugated anti-CD90 monoclonal antibody (Clone 5E10; eBioscience, San Diego, CA); an APC-conjugated anti-CD133/2 antibody (Clone 293C3; Miltenyi Biotec K.K.); an APC-conjugated anti-CD44 mouse monoclonal antibody (eBioscience); an APC-conjugated anti-CD13 antibody (eBioscience); and a PE-conjugated anti-CD24 antibody (Miltenyi Biotec K.K.).

Techniques: Marker, Expressing, Flow Cytometry, Labeling, FACS, Staining, Immunohistochemistry, Standard Deviation