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cd133 pe  (Miltenyi Biotec)


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    Structured Review

    Miltenyi Biotec cd133 pe
    Spheroid formation, maturation, and functional CSC enrichment in HGSOC cell lines cultured under ultra‐low attachment (ULA) conditions. Representative brightfield microscopy images of OVCAR‐3 and OVSAHO cell lines cultured in ULA plates at relevant time points (4× objective). Yellow arrows indicate highly compartmentalized OVCAR‐3 spheroids, whereas red arrows denote overcrowded aggregates observed at extended culture durations (a). Schematic illustration summarizing spheroid morphologies observed under ULA conditions, including loose aggregates, compact spheroids, and compartmentalized spheroids (b). Quantitative analysis of spheroid diameter distribution and representative size comparison of highly compartmentalized OVCAR‐3 spheroids illustrating intra‐well heterogeneity within ULA cultures at Day 48 (10× objective) (c). Representative confocal microscopy images of an OVCAR‐3 spheroid at Day 48 including brightfield image, confocal z‐sections acquired at ∼2 µm intervals, and corresponding z‐stack projection (63× objective). Blue = DAPI (nuclei), green = α‐tubulin. Individual channels are shown separately to facilitate visualization of marker <t>distribution.</t> <t>CD133‐PE</t> monolayer negative controls and uncropped confocal images are provided in Figure S1 (d). Representative confocal microscopy images of additional aggregates and spheroids from the same culture (63X objective). Blue = DAPI, green = α‐tubulin, red = CD133‐PE. Individual channels are shown separately. CD133‐PE monolayer negative controls and uncropped images are provided in Figure (e). Growth inhibition curves and corresponding IC50 bar graphs for carboplatin, niraparib, paclitaxel, olaparib, cisplatin, and doxorubicin comparing OVCAR‐3 monolayer cultures (black bars) and ULA spheroids (gray bars). Viability was assessed using the NCI‐SRB assay (f). Data are presented as mean ± SD from three independent biological replicates. For SRB assays, each biological replicate contained technical triplicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.
    Cd133 Pe, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd133+pe/CD133%2F1+Antibody%2C+anti-human%2C+REAfinity/pmc13420801-103-0-1
    Average 94 stars, based on 44 article reviews
    cd133 pe - by Bioz Stars, 2026-10
    94/100 stars

    Images

    1) Product Images from "Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer"

    Article Title: Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer

    Journal: Biotechnology Journal

    doi: 10.1002/biot.70287

    Spheroid formation, maturation, and functional CSC enrichment in HGSOC cell lines cultured under ultra‐low attachment (ULA) conditions. Representative brightfield microscopy images of OVCAR‐3 and OVSAHO cell lines cultured in ULA plates at relevant time points (4× objective). Yellow arrows indicate highly compartmentalized OVCAR‐3 spheroids, whereas red arrows denote overcrowded aggregates observed at extended culture durations (a). Schematic illustration summarizing spheroid morphologies observed under ULA conditions, including loose aggregates, compact spheroids, and compartmentalized spheroids (b). Quantitative analysis of spheroid diameter distribution and representative size comparison of highly compartmentalized OVCAR‐3 spheroids illustrating intra‐well heterogeneity within ULA cultures at Day 48 (10× objective) (c). Representative confocal microscopy images of an OVCAR‐3 spheroid at Day 48 including brightfield image, confocal z‐sections acquired at ∼2 µm intervals, and corresponding z‐stack projection (63× objective). Blue = DAPI (nuclei), green = α‐tubulin. Individual channels are shown separately to facilitate visualization of marker distribution. CD133‐PE monolayer negative controls and uncropped confocal images are provided in Figure S1 (d). Representative confocal microscopy images of additional aggregates and spheroids from the same culture (63X objective). Blue = DAPI, green = α‐tubulin, red = CD133‐PE. Individual channels are shown separately. CD133‐PE monolayer negative controls and uncropped images are provided in Figure (e). Growth inhibition curves and corresponding IC50 bar graphs for carboplatin, niraparib, paclitaxel, olaparib, cisplatin, and doxorubicin comparing OVCAR‐3 monolayer cultures (black bars) and ULA spheroids (gray bars). Viability was assessed using the NCI‐SRB assay (f). Data are presented as mean ± SD from three independent biological replicates. For SRB assays, each biological replicate contained technical triplicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.
    Figure Legend Snippet: Spheroid formation, maturation, and functional CSC enrichment in HGSOC cell lines cultured under ultra‐low attachment (ULA) conditions. Representative brightfield microscopy images of OVCAR‐3 and OVSAHO cell lines cultured in ULA plates at relevant time points (4× objective). Yellow arrows indicate highly compartmentalized OVCAR‐3 spheroids, whereas red arrows denote overcrowded aggregates observed at extended culture durations (a). Schematic illustration summarizing spheroid morphologies observed under ULA conditions, including loose aggregates, compact spheroids, and compartmentalized spheroids (b). Quantitative analysis of spheroid diameter distribution and representative size comparison of highly compartmentalized OVCAR‐3 spheroids illustrating intra‐well heterogeneity within ULA cultures at Day 48 (10× objective) (c). Representative confocal microscopy images of an OVCAR‐3 spheroid at Day 48 including brightfield image, confocal z‐sections acquired at ∼2 µm intervals, and corresponding z‐stack projection (63× objective). Blue = DAPI (nuclei), green = α‐tubulin. Individual channels are shown separately to facilitate visualization of marker distribution. CD133‐PE monolayer negative controls and uncropped confocal images are provided in Figure S1 (d). Representative confocal microscopy images of additional aggregates and spheroids from the same culture (63X objective). Blue = DAPI, green = α‐tubulin, red = CD133‐PE. Individual channels are shown separately. CD133‐PE monolayer negative controls and uncropped images are provided in Figure (e). Growth inhibition curves and corresponding IC50 bar graphs for carboplatin, niraparib, paclitaxel, olaparib, cisplatin, and doxorubicin comparing OVCAR‐3 monolayer cultures (black bars) and ULA spheroids (gray bars). Viability was assessed using the NCI‐SRB assay (f). Data are presented as mean ± SD from three independent biological replicates. For SRB assays, each biological replicate contained technical triplicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Techniques Used: Functional Assay, Cell Culture, Microscopy, Comparison, Confocal Microscopy, Marker, Inhibition, Sulforhodamine B Assay, Two Tailed Test

    Comparative benchmarking of CSC and EMT‐associated traits in HGSOC cells cultured using ULA and photopatterned GelMA platforms. Representative western blot images and corresponding quantitative bar graphs showing expression of CSC‐associated proteins SOX‐2, ALDH1A1, NANOG, and OCT‐4 in OVCAR‐3 and OVSAHO cultures. Protein levels were normalized to Vinculin and Calnexin, with representative loading control bands shown. Different loading controls were selected depending on target abundance, membrane compatibility, and subcellular localization. Quantification was performed using ImageJ (a). Representative flow cytometry pseudo‐dot plots with gates defined using unstained controls, and bar graphs summarizing the percentage of CD133‐PE, CD44‐FITC, CD117‐PE, and CXCR4‐FITC positive populations. Representative gating strategy is provided in Figure (b). Representative western blot images and quantitative bar graphs of EMT‐associated proteins Slug, Snail, TWIST1, vimentin, and fibronectin in OVCAR‐3 cultures (normalized to calnexin). E‐cadherin and N‐cadherin were additionally analyzed (normalized to α‐tubulin), with quantitative N‐cadherin/E‐cadherin ratios shown (c). Representative western blot analysis of stemness‐associated proteins in OVCAR‐3 cells cultured as 2D monolayer, 2D monolayer in spheroid enrichment medium, UV‐treated monolayer, ULA spheroids, and photopatterned GelMA (GelMA‐Pm) spheroids. Due to limited cell yield obtained from GelMA‐Pm cultures, only SOX‐2 and ALDH1A1 expression could be evaluated in these samples. Protein expression was normalized to GAPDH. Corresponding densitometric quantification is shown on the right. Full‐length uncropped blots are provided in Figure . (d). RT‐qPCR analysis of the stemness‐associated genes SOX‐2, NANOG, ALDH1A1, and c‐MYC in OVCAR‐3 cells cultured under 2D monolayer, 2D + sphere medium, 2D + UV, photopatterned GelMA (GelMA‐Pm), and ULA spheroid conditions. Gene expression levels were normalized to β‐actin and are presented relative to 2D monolayer controls (set to 1). Values above 1 indicate increased expression relative to monolayer cultures. Data represent mean ± SD from three independent biological replicates ( n = 3). Statistical significance was determined using multiple t ‐tests; significance symbols are defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001. Western blot images were cropped for clarity; non‐adjacent lanes from the same membrane are indicated by spaces. Full‐length blots are provided in Figure .
    Figure Legend Snippet: Comparative benchmarking of CSC and EMT‐associated traits in HGSOC cells cultured using ULA and photopatterned GelMA platforms. Representative western blot images and corresponding quantitative bar graphs showing expression of CSC‐associated proteins SOX‐2, ALDH1A1, NANOG, and OCT‐4 in OVCAR‐3 and OVSAHO cultures. Protein levels were normalized to Vinculin and Calnexin, with representative loading control bands shown. Different loading controls were selected depending on target abundance, membrane compatibility, and subcellular localization. Quantification was performed using ImageJ (a). Representative flow cytometry pseudo‐dot plots with gates defined using unstained controls, and bar graphs summarizing the percentage of CD133‐PE, CD44‐FITC, CD117‐PE, and CXCR4‐FITC positive populations. Representative gating strategy is provided in Figure (b). Representative western blot images and quantitative bar graphs of EMT‐associated proteins Slug, Snail, TWIST1, vimentin, and fibronectin in OVCAR‐3 cultures (normalized to calnexin). E‐cadherin and N‐cadherin were additionally analyzed (normalized to α‐tubulin), with quantitative N‐cadherin/E‐cadherin ratios shown (c). Representative western blot analysis of stemness‐associated proteins in OVCAR‐3 cells cultured as 2D monolayer, 2D monolayer in spheroid enrichment medium, UV‐treated monolayer, ULA spheroids, and photopatterned GelMA (GelMA‐Pm) spheroids. Due to limited cell yield obtained from GelMA‐Pm cultures, only SOX‐2 and ALDH1A1 expression could be evaluated in these samples. Protein expression was normalized to GAPDH. Corresponding densitometric quantification is shown on the right. Full‐length uncropped blots are provided in Figure . (d). RT‐qPCR analysis of the stemness‐associated genes SOX‐2, NANOG, ALDH1A1, and c‐MYC in OVCAR‐3 cells cultured under 2D monolayer, 2D + sphere medium, 2D + UV, photopatterned GelMA (GelMA‐Pm), and ULA spheroid conditions. Gene expression levels were normalized to β‐actin and are presented relative to 2D monolayer controls (set to 1). Values above 1 indicate increased expression relative to monolayer cultures. Data represent mean ± SD from three independent biological replicates ( n = 3). Statistical significance was determined using multiple t ‐tests; significance symbols are defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001. Western blot images were cropped for clarity; non‐adjacent lanes from the same membrane are indicated by spaces. Full‐length blots are provided in Figure .

    Techniques Used: Cell Culture, Western Blot, Expressing, Control, Membrane, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Two Tailed Test

    Optimization of photopatterned GelMA hydrogels for spheroid formation and CSC enrichment in HGSOC cell lines. Representative brightfield microscopy images of photopatterned GelMA (GelMA‐Pm) cultures of OVCAR‐3 and OVSAHO cells (upper panels: 4× objective; lower panels: 10× objective), showing spheroid formation under optimized conditions (a). Schematic overview of the GelMA‐Pm fabrication workflow and representative image of square photomask‐generated polymerized GelMA structures visible to the naked eye (b). Quantitative comparison of spheroid number and spheroid diameter per mm 2 between ULA and GelMA‐Pm cultures (c). Representative brightfield microscopy images illustrating approximate spheroid diameter distributions within photopatterned GelMA cultures at Day 14, including representative OVCAR‐3 spheroids (∼100 µm) and smaller OVSAHO spheroids (∼50 µm) (d). Representative confocal microscopy images of GelMA hydrogels containing spheroids from OVCAR‐3 and OVSAHO cultures, demonstrating cell viability and spatial distribution (10× objective, with selected regions magnified to 20X as indicated by yellow boxes). Blue = DAPI (nuclei), green = α‐tubulin, red = CD133‐PE. Corresponding brightfield and fluorescence images of identical fields are shown (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.
    Figure Legend Snippet: Optimization of photopatterned GelMA hydrogels for spheroid formation and CSC enrichment in HGSOC cell lines. Representative brightfield microscopy images of photopatterned GelMA (GelMA‐Pm) cultures of OVCAR‐3 and OVSAHO cells (upper panels: 4× objective; lower panels: 10× objective), showing spheroid formation under optimized conditions (a). Schematic overview of the GelMA‐Pm fabrication workflow and representative image of square photomask‐generated polymerized GelMA structures visible to the naked eye (b). Quantitative comparison of spheroid number and spheroid diameter per mm 2 between ULA and GelMA‐Pm cultures (c). Representative brightfield microscopy images illustrating approximate spheroid diameter distributions within photopatterned GelMA cultures at Day 14, including representative OVCAR‐3 spheroids (∼100 µm) and smaller OVSAHO spheroids (∼50 µm) (d). Representative confocal microscopy images of GelMA hydrogels containing spheroids from OVCAR‐3 and OVSAHO cultures, demonstrating cell viability and spatial distribution (10× objective, with selected regions magnified to 20X as indicated by yellow boxes). Blue = DAPI (nuclei), green = α‐tubulin, red = CD133‐PE. Corresponding brightfield and fluorescence images of identical fields are shown (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Techniques Used: Microscopy, Generated, Comparison, Confocal Microscopy, Fluorescence, Two Tailed Test

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    Incubation:

    Article Title: The magnitude of exercise‐induced progenitor cell mobilisation and extravasation is positively associated with cardiorespiratory fitness
    Article Snippet: .. Cell preparations were incubated with CD34‐FITC (BD Biosciences, cat. no. 345801, clone: 8G12, dilution: 1:20), CD45‐PerCP (BD Biosciences, cat. no.: 345809, clone: 2D1, dilution: 1:10), VEGFR2‐APC (R&D Systems, Minneapolis, MN, USA, cat. no.: FAB357A, clone: 89106, dilution: 1:30) CD133‐PE (Miltenyi Biotech, Bergisch Gladbach, Germany, cat. no.: 130‐080‐801, clone: AC133, dilution: 1:20) for 20 min in the dark at room temperature and subsequently washed in PBS by centrifuging at 250 g for 7 min. ..

    Article Title: Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer
    Article Snippet: .. CD133‐PE (Miltenyi Biotec, equal mixing of 130‐110‐962 and 130‐110‐962, 1:50–1:50 = 2:100) antibody was diluted 1:50 in CellO‐IF and incubation was done at 4°C for 20 min. ..

    Staining:

    Article Title: Pancreatic Cancer Stem Cells Co-Expressing SOX2, OCT4, and TERThigh Represent an Aggressive Subpopulation
    Article Snippet: Flow cytometry data were analyzed using the FlowJo version 10.8.1 software package (Tree Star, Ashland, OR, USA). .. For comparative phenotyping, a total of 4 × 105 sorted cells were stained with CD24PE (clone ML5, 555428, BD Biosciences, San Jose, CA, USA) and EpCAM-APC (347200), or cells were stained with CD133-PE (clone AC133, 130-112-157, Miltenyi Biotec, Bergisch Gladbach, Germany) and EpCAM-APC (clone HEA125, 347200, BD Biosciences, San Jose, CA, USA) for 30 min on ice. .. The isotypes IgG2a-PE (clone 130-117-787, Miltenyi Biotec, Bergisch Gladbach, Germany), IgG1-PE (clone 130-092-212, Miltenyi Biotec, Bergisch Gladbach, Germany), and IgG1-APC (clone 130-092-214, Miltenyi Biotec, Bergisch Gladbach, Germany) were used as the staining control.

    Article Title: Pancreatic Cancer Stem Cells Co-Expressing SOX2, OCT4, and TERT high Represent an Aggressive Subpopulation
    Article Snippet: Flow cytometry data were analyzed using the FlowJo version 10.8.1 software package (Tree Star, Ashland, OR, USA). .. For comparative phenotyping, a total of 4 × 10 5 sorted cells were stained with CD24-PE (clone ML5, 555428, BD Biosciences, San Jose, CA, USA) and EpCAM-APC (347200), or cells were stained with CD133-PE (clone AC133, 130-112-157, Miltenyi Biotec, Bergisch Gladbach, Germany) and EpCAM-APC (clone HEA125, 347200, BD Biosciences, San Jose, CA, USA) for 30 min on ice. .. The isotypes IgG2a-PE (clone 130-117-787, Miltenyi Biotec, Bergisch Gladbach, Germany), IgG1-PE (clone 130-092-212, Miltenyi Biotec, Bergisch Gladbach, Germany), and IgG1-APC (clone 130-092-214, Miltenyi Biotec, Bergisch Gladbach, Germany) were used as the staining control.

    Article Title: Targeted delivery of therapeutics to metastatic lung cancer cells using aptamer-conjugated nanoemulsions.
    Article Snippet: Lung cancer is the primary cause of cancer-related deaths, with a five-year overall survival rate of 15 %, mainly related to late diagnosis at advanced stages.. Despite recent improvements in cancer medicine, metastatic disease still, in fact, represents the main challenge for effective therapies.. Targeting disseminated cells requires a better understanding of the underlying biological mechanisms and a paradigm shift to devise innovative treatments.

    Flow Cytometry:

    Article Title: Erratum: Asiaticoside inhibits epithelial-mesenchymal transition and stem cell-like properties of pancreatic cancer PANC-1 cells by blocking the activation of p65 and p38MAPK
    Article Snippet: .. The CO2 incubator and automatic microplate reader were purchased from Thermo Scientific; flow cytometer was purchased from FASCAN Becton Dickison, USA; vernier calipers (precision 0.1 mm, EC-200) were purchased from Chengdu Measuring Tool & Cutting Tool Co., Ltd.; pancreatic cancer PANC-1 cells were purchased from the cell bank of the Shanghai Chinese Academy of Sciences; ATS was purchased from Guangxi Changzhou Natural Products Development Co., Ltd., with a purity of >95%; RPMI 1640 medium and mycoplasma-free fetal bovine serum were purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; CCK-8 detection kit and TUNEL tissue apoptosis detection kit were purchased from Shenyang Wanlei Biotechnology Co., Ltd.; β-actin monoclonal antibody was purchased from Beijing Soleibao Technology Co., Ltd.; CD133-PE and CD44-FITC antibodies were purchased from Miltenyi Biotec, Germany; rabbit anti-human E-cadherin, N-cadherin, vimentin, and p65, p-p65, p38 mitogen-activated protein kinase (p38MAPK), p-p38MAPK monoclonal antibodies, were purchased from Santa Cruz, USA; horseradish peroxidase (HRP) labeled goat anti-rabbit IgG was purchased from DAKO, Denmark. ..

    CCK-8 Assay:

    Article Title: Erratum: Asiaticoside inhibits epithelial-mesenchymal transition and stem cell-like properties of pancreatic cancer PANC-1 cells by blocking the activation of p65 and p38MAPK
    Article Snippet: .. The CO2 incubator and automatic microplate reader were purchased from Thermo Scientific; flow cytometer was purchased from FASCAN Becton Dickison, USA; vernier calipers (precision 0.1 mm, EC-200) were purchased from Chengdu Measuring Tool & Cutting Tool Co., Ltd.; pancreatic cancer PANC-1 cells were purchased from the cell bank of the Shanghai Chinese Academy of Sciences; ATS was purchased from Guangxi Changzhou Natural Products Development Co., Ltd., with a purity of >95%; RPMI 1640 medium and mycoplasma-free fetal bovine serum were purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; CCK-8 detection kit and TUNEL tissue apoptosis detection kit were purchased from Shenyang Wanlei Biotechnology Co., Ltd.; β-actin monoclonal antibody was purchased from Beijing Soleibao Technology Co., Ltd.; CD133-PE and CD44-FITC antibodies were purchased from Miltenyi Biotec, Germany; rabbit anti-human E-cadherin, N-cadherin, vimentin, and p65, p-p65, p38 mitogen-activated protein kinase (p38MAPK), p-p38MAPK monoclonal antibodies, were purchased from Santa Cruz, USA; horseradish peroxidase (HRP) labeled goat anti-rabbit IgG was purchased from DAKO, Denmark. ..

    TUNEL Assay:

    Article Title: Erratum: Asiaticoside inhibits epithelial-mesenchymal transition and stem cell-like properties of pancreatic cancer PANC-1 cells by blocking the activation of p65 and p38MAPK
    Article Snippet: .. The CO2 incubator and automatic microplate reader were purchased from Thermo Scientific; flow cytometer was purchased from FASCAN Becton Dickison, USA; vernier calipers (precision 0.1 mm, EC-200) were purchased from Chengdu Measuring Tool & Cutting Tool Co., Ltd.; pancreatic cancer PANC-1 cells were purchased from the cell bank of the Shanghai Chinese Academy of Sciences; ATS was purchased from Guangxi Changzhou Natural Products Development Co., Ltd., with a purity of >95%; RPMI 1640 medium and mycoplasma-free fetal bovine serum were purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; CCK-8 detection kit and TUNEL tissue apoptosis detection kit were purchased from Shenyang Wanlei Biotechnology Co., Ltd.; β-actin monoclonal antibody was purchased from Beijing Soleibao Technology Co., Ltd.; CD133-PE and CD44-FITC antibodies were purchased from Miltenyi Biotec, Germany; rabbit anti-human E-cadherin, N-cadherin, vimentin, and p65, p-p65, p38 mitogen-activated protein kinase (p38MAPK), p-p38MAPK monoclonal antibodies, were purchased from Santa Cruz, USA; horseradish peroxidase (HRP) labeled goat anti-rabbit IgG was purchased from DAKO, Denmark. ..

    Bioprocessing:

    Article Title: Erratum: Asiaticoside inhibits epithelial-mesenchymal transition and stem cell-like properties of pancreatic cancer PANC-1 cells by blocking the activation of p65 and p38MAPK
    Article Snippet: .. The CO2 incubator and automatic microplate reader were purchased from Thermo Scientific; flow cytometer was purchased from FASCAN Becton Dickison, USA; vernier calipers (precision 0.1 mm, EC-200) were purchased from Chengdu Measuring Tool & Cutting Tool Co., Ltd.; pancreatic cancer PANC-1 cells were purchased from the cell bank of the Shanghai Chinese Academy of Sciences; ATS was purchased from Guangxi Changzhou Natural Products Development Co., Ltd., with a purity of >95%; RPMI 1640 medium and mycoplasma-free fetal bovine serum were purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; CCK-8 detection kit and TUNEL tissue apoptosis detection kit were purchased from Shenyang Wanlei Biotechnology Co., Ltd.; β-actin monoclonal antibody was purchased from Beijing Soleibao Technology Co., Ltd.; CD133-PE and CD44-FITC antibodies were purchased from Miltenyi Biotec, Germany; rabbit anti-human E-cadherin, N-cadherin, vimentin, and p65, p-p65, p38 mitogen-activated protein kinase (p38MAPK), p-p38MAPK monoclonal antibodies, were purchased from Santa Cruz, USA; horseradish peroxidase (HRP) labeled goat anti-rabbit IgG was purchased from DAKO, Denmark. ..

    Labeling:

    Article Title: Erratum: Asiaticoside inhibits epithelial-mesenchymal transition and stem cell-like properties of pancreatic cancer PANC-1 cells by blocking the activation of p65 and p38MAPK
    Article Snippet: .. The CO2 incubator and automatic microplate reader were purchased from Thermo Scientific; flow cytometer was purchased from FASCAN Becton Dickison, USA; vernier calipers (precision 0.1 mm, EC-200) were purchased from Chengdu Measuring Tool & Cutting Tool Co., Ltd.; pancreatic cancer PANC-1 cells were purchased from the cell bank of the Shanghai Chinese Academy of Sciences; ATS was purchased from Guangxi Changzhou Natural Products Development Co., Ltd., with a purity of >95%; RPMI 1640 medium and mycoplasma-free fetal bovine serum were purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; CCK-8 detection kit and TUNEL tissue apoptosis detection kit were purchased from Shenyang Wanlei Biotechnology Co., Ltd.; β-actin monoclonal antibody was purchased from Beijing Soleibao Technology Co., Ltd.; CD133-PE and CD44-FITC antibodies were purchased from Miltenyi Biotec, Germany; rabbit anti-human E-cadherin, N-cadherin, vimentin, and p65, p-p65, p38 mitogen-activated protein kinase (p38MAPK), p-p38MAPK monoclonal antibodies, were purchased from Santa Cruz, USA; horseradish peroxidase (HRP) labeled goat anti-rabbit IgG was purchased from DAKO, Denmark. ..

    Immunopeptidomics:

    Article Title: Targeted delivery of therapeutics to metastatic lung cancer cells using aptamer-conjugated nanoemulsions.
    Article Snippet: Lung cancer is the primary cause of cancer-related deaths, with a five-year overall survival rate of 15 %, mainly related to late diagnosis at advanced stages.. Despite recent improvements in cancer medicine, metastatic disease still, in fact, represents the main challenge for effective therapies.. Targeting disseminated cells requires a better understanding of the underlying biological mechanisms and a paradigm shift to devise innovative treatments.

    Concentration Assay:

    Article Title: Identification and characterization of early human photoreceptor states and cell-state-specific retinoblastoma-related features
    Article Snippet: .. Dissociated cells were centrifuged and resuspended in a 5% fetal bovine serum in PBS (FBS/PBS) with CD133-PE (Miltenyi Biotec 130-113-108), CD44-FITC (BD Pharmingen 555478), and CD49b-FITC antibodies (BD Pharmingen 555498) to a final concentration of 10,000 cells/μl. ..



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    Image Search Results


    Spheroid formation, maturation, and functional CSC enrichment in HGSOC cell lines cultured under ultra‐low attachment (ULA) conditions. Representative brightfield microscopy images of OVCAR‐3 and OVSAHO cell lines cultured in ULA plates at relevant time points (4× objective). Yellow arrows indicate highly compartmentalized OVCAR‐3 spheroids, whereas red arrows denote overcrowded aggregates observed at extended culture durations (a). Schematic illustration summarizing spheroid morphologies observed under ULA conditions, including loose aggregates, compact spheroids, and compartmentalized spheroids (b). Quantitative analysis of spheroid diameter distribution and representative size comparison of highly compartmentalized OVCAR‐3 spheroids illustrating intra‐well heterogeneity within ULA cultures at Day 48 (10× objective) (c). Representative confocal microscopy images of an OVCAR‐3 spheroid at Day 48 including brightfield image, confocal z‐sections acquired at ∼2 µm intervals, and corresponding z‐stack projection (63× objective). Blue = DAPI (nuclei), green = α‐tubulin. Individual channels are shown separately to facilitate visualization of marker distribution. CD133‐PE monolayer negative controls and uncropped confocal images are provided in Figure S1 (d). Representative confocal microscopy images of additional aggregates and spheroids from the same culture (63X objective). Blue = DAPI, green = α‐tubulin, red = CD133‐PE. Individual channels are shown separately. CD133‐PE monolayer negative controls and uncropped images are provided in Figure (e). Growth inhibition curves and corresponding IC50 bar graphs for carboplatin, niraparib, paclitaxel, olaparib, cisplatin, and doxorubicin comparing OVCAR‐3 monolayer cultures (black bars) and ULA spheroids (gray bars). Viability was assessed using the NCI‐SRB assay (f). Data are presented as mean ± SD from three independent biological replicates. For SRB assays, each biological replicate contained technical triplicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Biotechnology Journal

    Article Title: Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer

    doi: 10.1002/biot.70287

    Figure Lengend Snippet: Spheroid formation, maturation, and functional CSC enrichment in HGSOC cell lines cultured under ultra‐low attachment (ULA) conditions. Representative brightfield microscopy images of OVCAR‐3 and OVSAHO cell lines cultured in ULA plates at relevant time points (4× objective). Yellow arrows indicate highly compartmentalized OVCAR‐3 spheroids, whereas red arrows denote overcrowded aggregates observed at extended culture durations (a). Schematic illustration summarizing spheroid morphologies observed under ULA conditions, including loose aggregates, compact spheroids, and compartmentalized spheroids (b). Quantitative analysis of spheroid diameter distribution and representative size comparison of highly compartmentalized OVCAR‐3 spheroids illustrating intra‐well heterogeneity within ULA cultures at Day 48 (10× objective) (c). Representative confocal microscopy images of an OVCAR‐3 spheroid at Day 48 including brightfield image, confocal z‐sections acquired at ∼2 µm intervals, and corresponding z‐stack projection (63× objective). Blue = DAPI (nuclei), green = α‐tubulin. Individual channels are shown separately to facilitate visualization of marker distribution. CD133‐PE monolayer negative controls and uncropped confocal images are provided in Figure S1 (d). Representative confocal microscopy images of additional aggregates and spheroids from the same culture (63X objective). Blue = DAPI, green = α‐tubulin, red = CD133‐PE. Individual channels are shown separately. CD133‐PE monolayer negative controls and uncropped images are provided in Figure (e). Growth inhibition curves and corresponding IC50 bar graphs for carboplatin, niraparib, paclitaxel, olaparib, cisplatin, and doxorubicin comparing OVCAR‐3 monolayer cultures (black bars) and ULA spheroids (gray bars). Viability was assessed using the NCI‐SRB assay (f). Data are presented as mean ± SD from three independent biological replicates. For SRB assays, each biological replicate contained technical triplicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: CD133‐PE (Miltenyi Biotec, equal mixing of 130‐110‐962 and 130‐110‐962, 1:50–1:50 = 2:100) antibody was diluted 1:50 in CellO‐IF and incubation was done at 4°C for 20 min.

    Techniques: Functional Assay, Cell Culture, Microscopy, Comparison, Confocal Microscopy, Marker, Inhibition, Sulforhodamine B Assay, Two Tailed Test

    Comparative benchmarking of CSC and EMT‐associated traits in HGSOC cells cultured using ULA and photopatterned GelMA platforms. Representative western blot images and corresponding quantitative bar graphs showing expression of CSC‐associated proteins SOX‐2, ALDH1A1, NANOG, and OCT‐4 in OVCAR‐3 and OVSAHO cultures. Protein levels were normalized to Vinculin and Calnexin, with representative loading control bands shown. Different loading controls were selected depending on target abundance, membrane compatibility, and subcellular localization. Quantification was performed using ImageJ (a). Representative flow cytometry pseudo‐dot plots with gates defined using unstained controls, and bar graphs summarizing the percentage of CD133‐PE, CD44‐FITC, CD117‐PE, and CXCR4‐FITC positive populations. Representative gating strategy is provided in Figure (b). Representative western blot images and quantitative bar graphs of EMT‐associated proteins Slug, Snail, TWIST1, vimentin, and fibronectin in OVCAR‐3 cultures (normalized to calnexin). E‐cadherin and N‐cadherin were additionally analyzed (normalized to α‐tubulin), with quantitative N‐cadherin/E‐cadherin ratios shown (c). Representative western blot analysis of stemness‐associated proteins in OVCAR‐3 cells cultured as 2D monolayer, 2D monolayer in spheroid enrichment medium, UV‐treated monolayer, ULA spheroids, and photopatterned GelMA (GelMA‐Pm) spheroids. Due to limited cell yield obtained from GelMA‐Pm cultures, only SOX‐2 and ALDH1A1 expression could be evaluated in these samples. Protein expression was normalized to GAPDH. Corresponding densitometric quantification is shown on the right. Full‐length uncropped blots are provided in Figure . (d). RT‐qPCR analysis of the stemness‐associated genes SOX‐2, NANOG, ALDH1A1, and c‐MYC in OVCAR‐3 cells cultured under 2D monolayer, 2D + sphere medium, 2D + UV, photopatterned GelMA (GelMA‐Pm), and ULA spheroid conditions. Gene expression levels were normalized to β‐actin and are presented relative to 2D monolayer controls (set to 1). Values above 1 indicate increased expression relative to monolayer cultures. Data represent mean ± SD from three independent biological replicates ( n = 3). Statistical significance was determined using multiple t ‐tests; significance symbols are defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001. Western blot images were cropped for clarity; non‐adjacent lanes from the same membrane are indicated by spaces. Full‐length blots are provided in Figure .

    Journal: Biotechnology Journal

    Article Title: Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer

    doi: 10.1002/biot.70287

    Figure Lengend Snippet: Comparative benchmarking of CSC and EMT‐associated traits in HGSOC cells cultured using ULA and photopatterned GelMA platforms. Representative western blot images and corresponding quantitative bar graphs showing expression of CSC‐associated proteins SOX‐2, ALDH1A1, NANOG, and OCT‐4 in OVCAR‐3 and OVSAHO cultures. Protein levels were normalized to Vinculin and Calnexin, with representative loading control bands shown. Different loading controls were selected depending on target abundance, membrane compatibility, and subcellular localization. Quantification was performed using ImageJ (a). Representative flow cytometry pseudo‐dot plots with gates defined using unstained controls, and bar graphs summarizing the percentage of CD133‐PE, CD44‐FITC, CD117‐PE, and CXCR4‐FITC positive populations. Representative gating strategy is provided in Figure (b). Representative western blot images and quantitative bar graphs of EMT‐associated proteins Slug, Snail, TWIST1, vimentin, and fibronectin in OVCAR‐3 cultures (normalized to calnexin). E‐cadherin and N‐cadherin were additionally analyzed (normalized to α‐tubulin), with quantitative N‐cadherin/E‐cadherin ratios shown (c). Representative western blot analysis of stemness‐associated proteins in OVCAR‐3 cells cultured as 2D monolayer, 2D monolayer in spheroid enrichment medium, UV‐treated monolayer, ULA spheroids, and photopatterned GelMA (GelMA‐Pm) spheroids. Due to limited cell yield obtained from GelMA‐Pm cultures, only SOX‐2 and ALDH1A1 expression could be evaluated in these samples. Protein expression was normalized to GAPDH. Corresponding densitometric quantification is shown on the right. Full‐length uncropped blots are provided in Figure . (d). RT‐qPCR analysis of the stemness‐associated genes SOX‐2, NANOG, ALDH1A1, and c‐MYC in OVCAR‐3 cells cultured under 2D monolayer, 2D + sphere medium, 2D + UV, photopatterned GelMA (GelMA‐Pm), and ULA spheroid conditions. Gene expression levels were normalized to β‐actin and are presented relative to 2D monolayer controls (set to 1). Values above 1 indicate increased expression relative to monolayer cultures. Data represent mean ± SD from three independent biological replicates ( n = 3). Statistical significance was determined using multiple t ‐tests; significance symbols are defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t ‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001. Western blot images were cropped for clarity; non‐adjacent lanes from the same membrane are indicated by spaces. Full‐length blots are provided in Figure .

    Article Snippet: CD133‐PE (Miltenyi Biotec, equal mixing of 130‐110‐962 and 130‐110‐962, 1:50–1:50 = 2:100) antibody was diluted 1:50 in CellO‐IF and incubation was done at 4°C for 20 min.

    Techniques: Cell Culture, Western Blot, Expressing, Control, Membrane, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Two Tailed Test

    Optimization of photopatterned GelMA hydrogels for spheroid formation and CSC enrichment in HGSOC cell lines. Representative brightfield microscopy images of photopatterned GelMA (GelMA‐Pm) cultures of OVCAR‐3 and OVSAHO cells (upper panels: 4× objective; lower panels: 10× objective), showing spheroid formation under optimized conditions (a). Schematic overview of the GelMA‐Pm fabrication workflow and representative image of square photomask‐generated polymerized GelMA structures visible to the naked eye (b). Quantitative comparison of spheroid number and spheroid diameter per mm 2 between ULA and GelMA‐Pm cultures (c). Representative brightfield microscopy images illustrating approximate spheroid diameter distributions within photopatterned GelMA cultures at Day 14, including representative OVCAR‐3 spheroids (∼100 µm) and smaller OVSAHO spheroids (∼50 µm) (d). Representative confocal microscopy images of GelMA hydrogels containing spheroids from OVCAR‐3 and OVSAHO cultures, demonstrating cell viability and spatial distribution (10× objective, with selected regions magnified to 20X as indicated by yellow boxes). Blue = DAPI (nuclei), green = α‐tubulin, red = CD133‐PE. Corresponding brightfield and fluorescence images of identical fields are shown (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Biotechnology Journal

    Article Title: Benchmarking Ultra‐Low Attachment and Photopatterned GelMA 3D Culture Platforms for Modeling Cancer Stemness in High‐Grade Serous Ovarian Cancer

    doi: 10.1002/biot.70287

    Figure Lengend Snippet: Optimization of photopatterned GelMA hydrogels for spheroid formation and CSC enrichment in HGSOC cell lines. Representative brightfield microscopy images of photopatterned GelMA (GelMA‐Pm) cultures of OVCAR‐3 and OVSAHO cells (upper panels: 4× objective; lower panels: 10× objective), showing spheroid formation under optimized conditions (a). Schematic overview of the GelMA‐Pm fabrication workflow and representative image of square photomask‐generated polymerized GelMA structures visible to the naked eye (b). Quantitative comparison of spheroid number and spheroid diameter per mm 2 between ULA and GelMA‐Pm cultures (c). Representative brightfield microscopy images illustrating approximate spheroid diameter distributions within photopatterned GelMA cultures at Day 14, including representative OVCAR‐3 spheroids (∼100 µm) and smaller OVSAHO spheroids (∼50 µm) (d). Representative confocal microscopy images of GelMA hydrogels containing spheroids from OVCAR‐3 and OVSAHO cultures, demonstrating cell viability and spatial distribution (10× objective, with selected regions magnified to 20X as indicated by yellow boxes). Blue = DAPI (nuclei), green = α‐tubulin, red = CD133‐PE. Corresponding brightfield and fluorescence images of identical fields are shown (e). Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined using unpaired two‐tailed Student's t‐tests or one‐way ANOVA where appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: CD133‐PE (Miltenyi Biotec, equal mixing of 130‐110‐962 and 130‐110‐962, 1:50–1:50 = 2:100) antibody was diluted 1:50 in CellO‐IF and incubation was done at 4°C for 20 min.

    Techniques: Microscopy, Generated, Comparison, Confocal Microscopy, Fluorescence, Two Tailed Test

    Positive and negative control cells for both marker panels. ( A ) HeLa (white arrows): in the first panel positive for cytokeratin (CK), Vimentin (Vim), VEGF and in the second panel positive for p16INK4A, negative for PD-L1 and CD45. ( B ) T98G (red arrows): positive for Vim. ( C ) CaSki (yellow arrows): positive for CK, Vim, VEGF, PD-L1 and p16INK4A. ( D ) MCF-7 (green arrows): positive for CK. All cell lines were negative for CD45. Only hematopoietic cells showed CD45 positive staining. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and the figure was assembled using Microsoft PowerPoint.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Positive and negative control cells for both marker panels. ( A ) HeLa (white arrows): in the first panel positive for cytokeratin (CK), Vimentin (Vim), VEGF and in the second panel positive for p16INK4A, negative for PD-L1 and CD45. ( B ) T98G (red arrows): positive for Vim. ( C ) CaSki (yellow arrows): positive for CK, Vim, VEGF, PD-L1 and p16INK4A. ( D ) MCF-7 (green arrows): positive for CK. All cell lines were negative for CD45. Only hematopoietic cells showed CD45 positive staining. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and the figure was assembled using Microsoft PowerPoint.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Negative Control, Marker, Staining, Software

    Representative images of patient-derived DTCs with three different profiles. ( A ) DTC positive for Vim, VEGF and p16, negative for CK, PD-L1 and CD45. ( B ) CK and Vim positive while negative for VEGF, PD-L1, p16 and CD45. ( C ) DTC positive for Vim and PD-L1, negative for all other markers. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and figure was assembled using Microsoft PowerPoint.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Representative images of patient-derived DTCs with three different profiles. ( A ) DTC positive for Vim, VEGF and p16, negative for CK, PD-L1 and CD45. ( B ) CK and Vim positive while negative for VEGF, PD-L1, p16 and CD45. ( C ) DTC positive for Vim and PD-L1, negative for all other markers. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and figure was assembled using Microsoft PowerPoint.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Derivative Assay, Software

    Comparison of p16, VEGF and PD-L1 in matching tumor tissue. In four cases we found VEGF on DTCs but not in matching tumor tissue, indicating a potential therapeutic target. Further, we detected PD-L1 on DTCs in 11 patients that had PD-L1 negative tissue at diagnosis, suggesting a potential discordance between tumor tissue and DTC phenotype. Chart generated using Microsoft Excel.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Comparison of p16, VEGF and PD-L1 in matching tumor tissue. In four cases we found VEGF on DTCs but not in matching tumor tissue, indicating a potential therapeutic target. Further, we detected PD-L1 on DTCs in 11 patients that had PD-L1 negative tissue at diagnosis, suggesting a potential discordance between tumor tissue and DTC phenotype. Chart generated using Microsoft Excel.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Comparison, Biomarker Discovery, Generated

    Study design. At diagnosis a tissue biopsy was obtained to histologically confirm cervical cancer cases. The FFPE tissue samples were stained against p16, PD-L1 and VEGF for comparison with DTC profiles. One day prior to oncologic surgery, blood samples were collected for CTC detection using the standardized CK based brightfield method. Bone marrow aspirates were sampled during surgery and DTCs were analyzed using the CK based and the multi-parameter IF staining method. Post-operative follow-up blood samples were collected within two years after surgery during routine visits if applicable. Clinical Outcome was assessed up to February 2025. Created with BioRender.com.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Study design. At diagnosis a tissue biopsy was obtained to histologically confirm cervical cancer cases. The FFPE tissue samples were stained against p16, PD-L1 and VEGF for comparison with DTC profiles. One day prior to oncologic surgery, blood samples were collected for CTC detection using the standardized CK based brightfield method. Bone marrow aspirates were sampled during surgery and DTCs were analyzed using the CK based and the multi-parameter IF staining method. Post-operative follow-up blood samples were collected within two years after surgery during routine visits if applicable. Clinical Outcome was assessed up to February 2025. Created with BioRender.com.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Biomarker Discovery, Staining, Comparison

    Sequential multi-parameter immunofluorescent staining. After applying the first antibody panel (CK, Vim and VEGF), the slides were scanned and DTC profiles were assessed. Subsequently, releasable fluorochrome-conjugates were digested and the slides were subjected to the second antibody panel (PD-L1, p16, CD45) followed by scanning and DTC detection. Created with BioRender.com.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Sequential multi-parameter immunofluorescent staining. After applying the first antibody panel (CK, Vim and VEGF), the slides were scanned and DTC profiles were assessed. Subsequently, releasable fluorochrome-conjugates were digested and the slides were subjected to the second antibody panel (PD-L1, p16, CD45) followed by scanning and DTC detection. Created with BioRender.com.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Staining

    Box plot showing the residual fluorescence signal after the release step (%) in 295 CaSki cells following treatment with a VEGF-PE conjugated antibody. The release efficiency was evaluated by signal quantification in the cells before and after treatment with the release reagent. The median fluorescence signal was 24.5% (mean 26.7%), corresponding to a signal reduction of 75.5%. The range was 1.75% to 87.78%. Statistical graph was generated using IBM SPSS Statistics Version 29.0.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: Box plot showing the residual fluorescence signal after the release step (%) in 295 CaSki cells following treatment with a VEGF-PE conjugated antibody. The release efficiency was evaluated by signal quantification in the cells before and after treatment with the release reagent. The median fluorescence signal was 24.5% (mean 26.7%), corresponding to a signal reduction of 75.5%. The range was 1.75% to 87.78%. Statistical graph was generated using IBM SPSS Statistics Version 29.0.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Fluorescence, Generated

    CaSki cells before and after the release step. Representative immunofluorescence image of two CaSki cells spiked into bone marrow cells ( A ) before and ( B ) after the release step. The PE channel shows VEGF staining before staining with CD45, while nuclei were stained with DAPI (blue). Images were processed using Zeiss ZEN software and figure was assembled using Microsoft PowerPoint.

    Journal: International Journal of Molecular Sciences

    Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

    doi: 10.3390/ijms27114875

    Figure Lengend Snippet: CaSki cells before and after the release step. Representative immunofluorescence image of two CaSki cells spiked into bone marrow cells ( A ) before and ( B ) after the release step. The PE channel shows VEGF staining before staining with CD45, while nuclei were stained with DAPI (blue). Images were processed using Zeiss ZEN software and figure was assembled using Microsoft PowerPoint.

    Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

    Techniques: Immunofluorescence, Staining, Software