anti cd133 monoclonal  (Novus Biologicals)

Journal: Photochemistry and photobiology

Article Title: Red light-activated depletion of drug-refractory glioblastoma stem cells and chemosensitization of an acquired-resistant mesenchymal phenotype

doi: 10.1111/php.13985

Figure Lengend Snippet: Patient-derived spheroid cultures of intrinsically chemosensitive and –resistant GSCs. ( a ) Simplified schematic of the process for establishing patient derived spheroid cultures enriched with GSCs from patient tumor biopsies adapted from Vescovi et al . 1 . ( b ) Bright field ( left ) and fluorescence confocal microscopy images of MGG6 spheroids stained with anti-CD133 antibody ( middle ) and with anti-nestin antibody ( right ). ( c ) Representative flow cytometry profiles of dissociated MGG6, MGG8 and MGG23 spheroids stained with anti-CD133 antibody labeled with phycoerythrin (PE) and anti-CD44 antibody labeled with Alexa Fluor 647 (Alexa 647) ( n = 3 biological replicates per condition). ( d ) Temozolomide dose response of the three GSC spheroid cultures with the green box indicating the peak clinical dose of temozolomide (5–50 μM) 48 , 49 and the solid trend lines resulting from a dose-response inhibitor fitting function ( Methods ); and, ( e ) estimated temozolomide EC50 doses for the three spheroid cultures resulting from the fitting function in d . Results are mean ± s.e.m ( n ≥ 3 biological replicates per condition; **** P < 0.001, one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Live cell staining of cell-surface CD133 was performed in the culture medium using an anti-human CD133 monoclonal antibody conjugated to R-phycoerythrin (CD133/2, clone 293C3; Miltenyi Biotec) at 37°C.

Techniques: Derivative Assay, Fluorescence, Confocal Microscopy, Staining, Flow Cytometry, Labeling

Journal: Photochemistry and photobiology

Article Title: Red light-activated depletion of drug-refractory glioblastoma stem cells and chemosensitization of an acquired-resistant mesenchymal phenotype

doi: 10.1111/php.13985

Figure Lengend Snippet: Patient-specific spheroid culture responses to temozolomide cycling with rapid acquisition of chemoresistance coinciding with enrichment of Mes– and PN/Mes–GSCs in MGG8 spheroids. ( a ) Schematic of weekly temozolomide treatment cycles with color coding of the GSC subtypes adapted from Brown et al . 44 . ( b ) CD133, CD44 and GSC subtype population profiles resulting from temozolomide cycling measured by flow cytometry for MGG6, MGG8 and MGG23 spheroid cultures. Results are mean ± s.e.m. Asterisks denote significance compared with the no-treatment (NT) control ( n = 3 biological replicates per condition, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test for MGG6 and MGG8 and an unpaired, two-tailed t -test for MGG23). ( c ) Temozolomide dose response for NT, TMZ+3 and TMZ+6 MGG8 spheroids with the green box indicating the peak clinical dose of temozolomide (5–50 μM) 48 , 49 and the solid trend lines resulting from a dose-response inhibitor fitting function ( Methods ). ( d ) Estimated temozolomide EC50 doses for each condition resulting from the fitting function in c . Results are mean ± s.e.m. The asterisks in ( d ) denote significance compared with untreated spheroids and among the TMZ-cycled groups ( n = 3 biological replicates per condition, *** P < 0.001, **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Live cell staining of cell-surface CD133 was performed in the culture medium using an anti-human CD133 monoclonal antibody conjugated to R-phycoerythrin (CD133/2, clone 293C3; Miltenyi Biotec) at 37°C.

Techniques: Flow Cytometry, Control, Two Tailed Test

Journal: Photochemistry and photobiology

Article Title: Red light-activated depletion of drug-refractory glioblastoma stem cells and chemosensitization of an acquired-resistant mesenchymal phenotype

doi: 10.1111/php.13985

Figure Lengend Snippet: Red light-activated therapy induces death in all GSC subtypes at clinically relevant doses. ( a ) All GSC subtypes convert non-fluorescent ALA into fluorescent and photodynamic PpIX (0.5 mM ALA for 24 h). Mean histograms of PpIX fluorescence of the four GSC subtype populations (CD133−/CD44 low , CD133−/CD44 high , CD133+/CD44 low and CD133+/CD44 high ) for the three patient-derived spheroid cultures analyzed by flow cytometry ( n = 3 biological replicates per condition). The control for each spheroid culture indicates cell autofluorescence background in the absence of ALA. ( b ) All three ALA-treated GSC spheroid cultures respond to light-activated therapy (635 nm) over a clinically relevant dose range (0.5 mM ALA for 24 h; 2–20 J/cm 2 ). Solid trend lines result from applying a dose-response inhibitor fitting function ( Methods ). ( c ) Estimated EC50 light doses for each of the three GSC spheroid cultures resulting from the dose-response fits shown in b . Results are mean ± s.e.m. ( n = 3 biological replicates per condition, ** P < 0.01, **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Live cell staining of cell-surface CD133 was performed in the culture medium using an anti-human CD133 monoclonal antibody conjugated to R-phycoerythrin (CD133/2, clone 293C3; Miltenyi Biotec) at 37°C.

Techniques: Fluorescence, Derivative Assay, Flow Cytometry, Control

Journal: Photochemistry and photobiology

Article Title: Red light-activated depletion of drug-refractory glioblastoma stem cells and chemosensitization of an acquired-resistant mesenchymal phenotype

doi: 10.1111/php.13985

Figure Lengend Snippet: Low-dose red light-activation of ALA-treated spheroid cultures sensitizes MGMT-negative GSCs with acquired drug-resistance to temozolomide. ( a ) Light dose response of MGG8 TMZ+6 cells evaluated 72 h post red irradiation. Results are mean ± s.e.m. Asterisks denote significance compared with the no-treatment (NT) control ( n = 3 biological replicates per condition; *** P < 0.001, **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test). Light only without ALA (10 J/cm 2 ; LO) and ALA without light (0 J/cm 2 ; ALA only) controls demonstrate minimal cell death compared to red light-activated treatment (ALA with light; 1, 5 and 10 J/cm 2 ). ( b ) Change in CD133 and CD44 cell-surface expression profiles of MGG8-TMZ+6 cells in response to different doses of PDT ( i.e. , varying doses of red light). Results are mean ± s.e.m. Asterisks denote significance compared with the no-treatment (NT) control ( n = 2 biological replicates per condition; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test). ( c ) Temozolomide dose response curve for no-treatment control (NT), TMZ+3, TMZ+6 and PDT treated (5 J/cm 2 ) TMZ+6 MGG8 spheroids; and, ( d ) estimated EC50 doses of temozolomide from analysis of the dose response curves shown in c using a dose-response inhibitor fitting function ( Methods ). MGG8-TMZ+6 spheroids became re-sensitized to temozolomide treatment after a single, sub-lethal light dose. Results are mean ± s.e.m. ( n = 3 biological replicates per condition; **** P < 0.0001, one-way ANOVA with Tukey’s post hoc test). ( e ) Temozolomide dose response of NT and red light-activation of ALA-treated MGMT-positive MGG23 spheroids; and, ( f ) estimated EC50 temozolomide doses for NT and light-activation of ALA-treated MGG23 spheroids. Light-activation did not induce sensitivity to temozolomide in these intrinsically drug-resistant spheroids (no significant difference was observed). Results are mean ± s.e.m. ( n = 7 biological replicates per condition; unpaired, two-tailed t -test).

Article Snippet: Live cell staining of cell-surface CD133 was performed in the culture medium using an anti-human CD133 monoclonal antibody conjugated to R-phycoerythrin (CD133/2, clone 293C3; Miltenyi Biotec) at 37°C.

Techniques: Activation Assay, Irradiation, Control, Expressing, Two Tailed Test

Journal: Turkish Journal of Surgery

Article Title: Co-expression of Stem Cell Markers CD133 and CD44 as Predictors of Metastatic Potential of Colorectal Carcinoma

doi: 10.47717/turkjsurg.2025.6837

Figure Lengend Snippet: Frequency of high and low co-expression of CD133/CD44 in groups of patients according to stage of disease.

Article Snippet: The primary antibodies used were anti-CD133 rabbit monoclonal antibody (Miltenyi, Germany; clone AC133, dilution 1:11) and anti-CD44 mouse monoclonal antibody (Novocastra, UK; clone DF1485, dilution 1:50).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Substituents introduction of methyl and methoxy functional groups on resveratrol stabilizes mTOR binding for autophagic cell death induction

doi: 10.1038/s41598-025-98616-6

Figure Lengend Snippet: Suppressive effects of SM-3 on CSCs markers via mTOR inhibition during spheroids and organoids formation in A549 cells. ( A ) The A549 cells were treated with SM-3 (50 µM) and observed using a phase-contrast microscope after 3 and 7 days of incubated primary and secondary spheroids. The relative number of spheroids were analyzed using ImageJ software. Scale bar: 100 μm (magnification: 4x) ( B ) Single spheroids from a CSCs-rich population of A549 cells were treated with SM-3 (50 µM) for 3 days, and CSCs viability was evaluated using Hoechst 33,342/PI double staining. Scale bar: 50 μm (magnification: 20x) ( C , D ) The CSCs-rich spheroids were treated with SM-3 (50 µM) for 24 h. The levels of upstream proteins mTOR and pAkt in SM-3 treated CSCs-rich spheroids were assessed using immunofluorescence. The SM-3 treated A549 spheroids were examined for the expression of stem cell markers, as well as the transcription factors. Scale bar: 50 μm (magnification: 20x) ( E ) SM-3 targeted CSCs by inhibiting mTOR during 3D organoids formation in A549 cells, as analyzed by immunofluorescence. The expression levels of pAkt (Ser473), mTOR, CSCs-rich transcription factors (CD133 and CD44), and stem cell markers (OCT4 and SOX2) were measured in SM-3-treated organoids. Scale bar: 20 μm (magnification: 40x) by confocal microscope ( F ) The fluorescence intensity was measured by image J software. ( G ) Cells were treated with SM-3 (50 µM), and the mRNA expression levels of the stem cell transcription factors OCT4, NANOG, and SOX2 were measured. The mRNA levels were normalized against the housekeeping gene GAPDH, and relative mRNA expression was calculated using comparative Ct cycles. ( H ) The heat map displays the fluorescence intensity of stem cell markers and stem cell transcription factors analyzed by using Image J software. Scale bar: 10 μm (magnification: 40x) (I) The protein expression levels of stem cell markers were assessed using western blot analysis, with the blot reprobed for β-actin to ensure equal protein loading. The parent compound Res (50 µM) served as the positive control. Data are presented as mean ± SD ( n = 3). Significance is indicated as * p < 0.05, ** p < 0.01, *** p < 0.001 compared to untreated control cells, and # p < 0.05, ## p < 0.01, ### p < 0.001 compared to Res-treated A549 cells.

Article Snippet: The mouse monoclonal antibodies for CD133 (cat no: MAB4399-I) from Millipore sigma.

Techniques: Inhibition, Microscopy, Incubation, Software, Double Staining, Immunofluorescence, Expressing, Fluorescence, Western Blot, Positive Control, Control