Structured Review

Greiner Bio t25 flasks
Detection of Venus-fluorescence and A . t .-sequences in recipient cells after passaging. (a) 2x10 5 hMSC were seeded into <t>T25,</t> incubated overnight to reach adherence (d0) and fed with EV derived from A . t .-hMSC cultures for 2 weeks. (b) Venus-positive cells were detected in 2 of 4 flasks (d14). One flask with 7 positive cells was passaged into 4xT25 flasks. (c) 7 days later (d21), one flask contained 10 Venus-positive cells. This culture was expanded again into 4xT25. (d) Venus-positive cells at d28 were evident in 2 flasks out of 4 with 13 cells in one flask and 1 cell in the second flask. Exemplarily, one positive MSC spot with corresponding phase-contrast for each time point is shown. Magnification x 100. (e, f) DNA of the flask with 13 Venus-positive cells was pretested in nested SYBR Green-based qPCR. Out of 10 primary reaction tubes, 4 were positive in the nested qPCR tested in 8 replicates (tubes 2, 4, 7 and 8; not shown) and were retested in TaqMan-based qPCR (e) and ddPCR (f). Shown are the results for positive control (pc, 10 copies/PCR reaction; 4 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), negative control (nc, untransduced hMSC; 8 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), and tube 2 and 4 (16 replicates in TaqMan-based qPCR and ddPCR).
T25 Flasks, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles"

Article Title: Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles

Journal: PLoS ONE

doi: 10.1371/journal.pone.0163665

Detection of Venus-fluorescence and A . t .-sequences in recipient cells after passaging. (a) 2x10 5 hMSC were seeded into T25, incubated overnight to reach adherence (d0) and fed with EV derived from A . t .-hMSC cultures for 2 weeks. (b) Venus-positive cells were detected in 2 of 4 flasks (d14). One flask with 7 positive cells was passaged into 4xT25 flasks. (c) 7 days later (d21), one flask contained 10 Venus-positive cells. This culture was expanded again into 4xT25. (d) Venus-positive cells at d28 were evident in 2 flasks out of 4 with 13 cells in one flask and 1 cell in the second flask. Exemplarily, one positive MSC spot with corresponding phase-contrast for each time point is shown. Magnification x 100. (e, f) DNA of the flask with 13 Venus-positive cells was pretested in nested SYBR Green-based qPCR. Out of 10 primary reaction tubes, 4 were positive in the nested qPCR tested in 8 replicates (tubes 2, 4, 7 and 8; not shown) and were retested in TaqMan-based qPCR (e) and ddPCR (f). Shown are the results for positive control (pc, 10 copies/PCR reaction; 4 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), negative control (nc, untransduced hMSC; 8 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), and tube 2 and 4 (16 replicates in TaqMan-based qPCR and ddPCR).
Figure Legend Snippet: Detection of Venus-fluorescence and A . t .-sequences in recipient cells after passaging. (a) 2x10 5 hMSC were seeded into T25, incubated overnight to reach adherence (d0) and fed with EV derived from A . t .-hMSC cultures for 2 weeks. (b) Venus-positive cells were detected in 2 of 4 flasks (d14). One flask with 7 positive cells was passaged into 4xT25 flasks. (c) 7 days later (d21), one flask contained 10 Venus-positive cells. This culture was expanded again into 4xT25. (d) Venus-positive cells at d28 were evident in 2 flasks out of 4 with 13 cells in one flask and 1 cell in the second flask. Exemplarily, one positive MSC spot with corresponding phase-contrast for each time point is shown. Magnification x 100. (e, f) DNA of the flask with 13 Venus-positive cells was pretested in nested SYBR Green-based qPCR. Out of 10 primary reaction tubes, 4 were positive in the nested qPCR tested in 8 replicates (tubes 2, 4, 7 and 8; not shown) and were retested in TaqMan-based qPCR (e) and ddPCR (f). Shown are the results for positive control (pc, 10 copies/PCR reaction; 4 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), negative control (nc, untransduced hMSC; 8 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), and tube 2 and 4 (16 replicates in TaqMan-based qPCR and ddPCR).

Techniques Used: Fluorescence, Passaging, Incubation, Derivative Assay, SYBR Green Assay, Real-time Polymerase Chain Reaction, Positive Control, Polymerase Chain Reaction, Negative Control

2) Product Images from "Electron Scattering in Conventional Cell Flask Experiments and Dose Distribution Dependency"

Article Title: Electron Scattering in Conventional Cell Flask Experiments and Dose Distribution Dependency

Journal: Scientific Reports

doi: 10.1038/s41598-019-57029-y

Analysis of colony distributions after 12 Gy at CSU. ( A ) Uneven colony distribution with increased survival near the wall of the flasks for 4 MeV compared to 18 MeV electron irradiation. 100,000 cells were plated to T25 flasks, T75 flask and glass dishes. ( B ) Increased colony survival near the edge of the 6 cm plastic dish after 25 J/m 2 of UV-C exposure. ( C ) Quantitative analysis of colony distribution using 4, 9, and 18 MeV. 100,000 cells were plated. Error bars indicate the standard error of the means for three independent experiments. Note the increased number of surviving colonies near the edges of the flask at 4 MeV.
Figure Legend Snippet: Analysis of colony distributions after 12 Gy at CSU. ( A ) Uneven colony distribution with increased survival near the wall of the flasks for 4 MeV compared to 18 MeV electron irradiation. 100,000 cells were plated to T25 flasks, T75 flask and glass dishes. ( B ) Increased colony survival near the edge of the 6 cm plastic dish after 25 J/m 2 of UV-C exposure. ( C ) Quantitative analysis of colony distribution using 4, 9, and 18 MeV. 100,000 cells were plated. Error bars indicate the standard error of the means for three independent experiments. Note the increased number of surviving colonies near the edges of the flask at 4 MeV.

Techniques Used: Irradiation

Related Articles

Transduction:

Article Title: Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles
Article Snippet: .. EV-transfer to recipient cells Recipient human BM-hMSC (2.5-5x104 ) unrelated to hMSC used for A .t .-transduction were seeded into T25-flasks (Greiner). .. Single, not pooled EV preparations derived from three individual A .t .-hMSC cultures were divided into two parts.

Clone Assay:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Ten shRNA knockdown sequences and one non-targeting sequence [ ] were conceived using RNAi Block-IT Designer (Invitrogen, Carlsbad, CA) and cloned into plasmids using standard techniques. .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

Cytometry:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1. .. All treatment groups were removed from culture flasks 48-h post-transfection for flow cytometry.

Blocking Assay:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Ten shRNA knockdown sequences and one non-targeting sequence [ ] were conceived using RNAi Block-IT Designer (Invitrogen, Carlsbad, CA) and cloned into plasmids using standard techniques. .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

Ex Vivo:

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: We adopted methods from evidence that inhibition of TGF-ß1 signaling corrected abnormal COL1A1 -to-COL1A2 gene expression ratios and increased alizarin red staining [ ] to explore whether similar intervention could influence expression of the signature gene COL1A2 and restore the ex vivo mineralization function to donor#6 cells. .. Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction.

Incubation:

Article Title: Cellular uptake, genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells
Article Snippet: .. The cells were seeded at a concentration of 2.5 × 105 cells per mL and incubated at 37 °C under CO2 atmosphere (5% CO2 in air) for 24 h in T25 flasks (Greiner Bio-one), and were treated with bare or silica-coated MNPs at a concentration range of 15–500 μg mL–1 for 4 and 24 h. The complete medium and mitomycin C (MMC; Sigma-Aldrich, USA; 0.6 μg mL–1 for 4 h and 0.3 μg mL–1 for 24 h) were used as negative and positive controls, respectively. .. At the end of the treatments, the cells were washed with complete medium, and cytochalasin B (Sigma-Aldrich, USA; final concentration of 6 μg mL–1 ) was added in the last 24 h of the culture.

Article Title: Subcellular Distribution of S-Nitrosylated H-Ras in Differentiated and Undifferentiated PC12 Cells during Hypoxia
Article Snippet: PC12 cell line In our experimental study, we cultured PC12 (Adh; ATCC® CRL1721.1™) cells in T25 flasks (Greiner Bio-One GmbH, Cat. No.: 690 170) in a humidified atmosphere that contained 5% CO 2 at 37˚C in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, ATCC® 30-2002™) supplemented with 10% heat- inactivated horse serum (Sigma-Aldrich), 5% fetal bovine serum (FBS, Sigma-Aldrich), 100 IU/ml penicillin and 50 µg/ml gentamycin sulphate. .. For the induction of differentiation, PC12 cells were incubated in low serum-DMEM that consisted of 1% heat-inactivated horse serum and 1% FBS, supplemented with nerve growth factor (NGF) 100 ng/ml for 5 days.

Article Title: Binding of Integrin ?6?4 to Plectin Prevents Plectin Association with F-Actin but Does Not Interfere with Intermediate Filament Binding
Article Snippet: The tissue was cut into several pieces and incubated in 0.5% (wt/vol) trypsin, 0.1% (wt/vol) EDTA for 90 min at 37°C to separate the epidermis from the dermis. .. The keratinocyte suspension was plated into two T25 flasks (Greiner) together with 5 × 105 irradiated 3T3 feeder cells and keratinocyte medium ( ) without EGF.

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction. .. All culture vessels were incubated at 37°C with 5% CO2 in a humidified incubator.

Expressing:

Article Title: Imatinib mesylate, a new kid on the block for the treatment of anti-neutrophil cytoplasmic autoantibodies-associated vasculitis?
Article Snippet: The cells were seeded in T25 flasks (Greiner, Frickenhausen, Germany) coated with gelatin (1%) and cultured in endothelial cell growth medium (EGM) (Promocell, Heidelberg, Germany). .. Characterization of endothelial cells was performed on the basis of a positive uptake of acetylated low-density lipoprotein, Factor VIII-related antigen and platelet/endothelial cell adhesion molecule 1 (PECAM) (CD31) expression, and a negative staining for alpha smooth muscle actin.

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: The selected plasmid allowed simultaneous expression of shRNA via the U6 promoter and CMV-driven enhanced green fluorescent protein (GFP) expression [ ] and is intended for making single-stranded AAV vectors. .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: We adopted methods from evidence that inhibition of TGF-ß1 signaling corrected abnormal COL1A1 -to-COL1A2 gene expression ratios and increased alizarin red staining [ ] to explore whether similar intervention could influence expression of the signature gene COL1A2 and restore the ex vivo mineralization function to donor#6 cells. .. Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction.

Modification:

Article Title: Electron Scattering in Conventional Cell Flask Experiments and Dose Distribution Dependency
Article Snippet: Cell culture Chinese hamster ovary (CHO) wild type CHO10B2 was kindly provided by Dr. Joel Bedford at CSU and maintained at 37 °C in a 5% CO2 incubator in Eagle’s minimal essential medium with alpha modification supplemented with 10% heat-inactivated fetal bovine serum, penicillin and streptomycin. .. For geometric analysis, T25 flasks marked with a measuring grid spacing of 4 mm (Greiner Bio-One, Monroe, NC) were used (Fig. ).

Article Title: Subcellular Distribution of S-Nitrosylated H-Ras in Differentiated and Undifferentiated PC12 Cells during Hypoxia
Article Snippet: .. PC12 cell line In our experimental study, we cultured PC12 (Adh; ATCC® CRL1721.1™) cells in T25 flasks (Greiner Bio-One GmbH, Cat. No.: 690 170) in a humidified atmosphere that contained 5% CO 2 at 37˚C in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, ATCC® 30-2002™) supplemented with 10% heat- inactivated horse serum (Sigma-Aldrich), 5% fetal bovine serum (FBS, Sigma-Aldrich), 100 IU/ml penicillin and 50 µg/ml gentamycin sulphate. .. For the induction of differentiation, PC12 cells were incubated in low serum-DMEM that consisted of 1% heat-inactivated horse serum and 1% FBS, supplemented with nerve growth factor (NGF) 100 ng/ml for 5 days.

Derivative Assay:

Article Title: Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles
Article Snippet: EV-transfer to recipient cells Recipient human BM-hMSC (2.5-5x104 ) unrelated to hMSC used for A .t .-transduction were seeded into T25-flasks (Greiner). .. Single, not pooled EV preparations derived from three individual A .t .-hMSC cultures were divided into two parts.

Transfection:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1. .. Media containing transfection complexes were removed 24-h post-exposure; new non-supplemented Minimum Essential Media, Eagle (MEM-E) (Mediatech, Herndon, VA) was added, either with or without 1 μg/ml LPS ( Escherichia coli O111:B4; Sigma–Aldrich, St. Louis, MO) for an additional 24-h.

Biomarker Assay:

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: Paragraph title: Biomarker signature gene verification; a role in outlier phenotypes ... Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction.

Cell Culture:

Article Title: Imatinib mesylate, a new kid on the block for the treatment of anti-neutrophil cytoplasmic autoantibodies-associated vasculitis?
Article Snippet: .. The cells were seeded in T25 flasks (Greiner, Frickenhausen, Germany) coated with gelatin (1%) and cultured in endothelial cell growth medium (EGM) (Promocell, Heidelberg, Germany). .. Confluent monolayers were passaged by Trypsin/ethylenediamine tetraacetic acid (EDTA) (Sigma-Aldrich).

Article Title: Electron Scattering in Conventional Cell Flask Experiments and Dose Distribution Dependency
Article Snippet: Paragraph title: Cell culture ... For geometric analysis, T25 flasks marked with a measuring grid spacing of 4 mm (Greiner Bio-One, Monroe, NC) were used (Fig. ).

Article Title: Subcellular Distribution of S-Nitrosylated H-Ras in Differentiated and Undifferentiated PC12 Cells during Hypoxia
Article Snippet: .. PC12 cell line In our experimental study, we cultured PC12 (Adh; ATCC® CRL1721.1™) cells in T25 flasks (Greiner Bio-One GmbH, Cat. No.: 690 170) in a humidified atmosphere that contained 5% CO 2 at 37˚C in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, ATCC® 30-2002™) supplemented with 10% heat- inactivated horse serum (Sigma-Aldrich), 5% fetal bovine serum (FBS, Sigma-Aldrich), 100 IU/ml penicillin and 50 µg/ml gentamycin sulphate. .. For the induction of differentiation, PC12 cells were incubated in low serum-DMEM that consisted of 1% heat-inactivated horse serum and 1% FBS, supplemented with nerve growth factor (NGF) 100 ng/ml for 5 days.

Article Title: Constitutively elevated levels of SOCS1 suppress innate responses in DF-1 immortalised chicken fibroblast cells
Article Snippet: .. They were seeded in T25 flasks (Greiner Bio One; 5.6 × 106 cells/flask) and cultured overnight in 5.5 ml 199 media (Gibco, Invitrogen) supplemented with 8% heat-inactivated newborn bovine serum (NBCS; Gibco, Invitrogen), 10% tryptose phosphate broth (TPB; Sigma), 2% nystatin (Sigma) and 0.1% penicillin and streptomycin (Gibco, Invitrogen). .. DF-1 were propagated in Dulbecco’s minimal essential medium (DMEM) (Life Technologies) supplemented with 10% heat-inactivated fetal bovine serum (Life Technologies) and penicillin and streptomycin.

Article Title: A Novel Cell Force Sensor for Quantification of Traction during Cell Spreading and Contact Guidance
Article Snippet: .. They were routinely cultured in T25 flasks (Greiner, Bio-One North America Inc., Monroe, NC; 1:2) in a standard tissue culture incubator at 37°C, 100% humidity, and 5% CO2 . .. Medium M199 (Gibco BRL, Gaithersburg, MD) was supplemented with 25 mM HEPES, 20% fetal calf serum (PAA Laboratories GmbH, Pasching, Austria), 2 mM L-glutamine, penicillin, streptomycin, 0.15 mg/ml ECGF, and 5000 IU/ml heparin (Sigma, St. Louis, MO).

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1. .. Media containing transfection complexes were removed 24-h post-exposure; new non-supplemented Minimum Essential Media, Eagle (MEM-E) (Mediatech, Herndon, VA) was added, either with or without 1 μg/ml LPS ( Escherichia coli O111:B4; Sigma–Aldrich, St. Louis, MO) for an additional 24-h.

Article Title: Irradiation affects cellular properties and Eph receptor expression in human melanoma cells
Article Snippet: .. For growth assays, cells were precultured in T25-flasks (Greiner Bio-one), irradiated with 5 or 10 Gy and further cultured for another 6 d. Cells were plated with 1 × 105 cells per well in a 6-well plate. ..

Inhibition:

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: We adopted methods from evidence that inhibition of TGF-ß1 signaling corrected abnormal COL1A1 -to-COL1A2 gene expression ratios and increased alizarin red staining [ ] to explore whether similar intervention could influence expression of the signature gene COL1A2 and restore the ex vivo mineralization function to donor#6 cells. .. Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction.

Sequencing:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Ten shRNA knockdown sequences and one non-targeting sequence [ ] were conceived using RNAi Block-IT Designer (Invitrogen, Carlsbad, CA) and cloned into plasmids using standard techniques. .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

Negative Staining:

Article Title: Imatinib mesylate, a new kid on the block for the treatment of anti-neutrophil cytoplasmic autoantibodies-associated vasculitis?
Article Snippet: The cells were seeded in T25 flasks (Greiner, Frickenhausen, Germany) coated with gelatin (1%) and cultured in endothelial cell growth medium (EGM) (Promocell, Heidelberg, Germany). .. Characterization of endothelial cells was performed on the basis of a positive uptake of acetylated low-density lipoprotein, Factor VIII-related antigen and platelet/endothelial cell adhesion molecule 1 (PECAM) (CD31) expression, and a negative staining for alpha smooth muscle actin.

Isolation:

Article Title: Imatinib mesylate, a new kid on the block for the treatment of anti-neutrophil cytoplasmic autoantibodies-associated vasculitis?
Article Snippet: Human umbilical vein endothelial cells (HUVEC) were isolated from fresh umbilical cords as described previously [ ]. .. The cells were seeded in T25 flasks (Greiner, Frickenhausen, Germany) coated with gelatin (1%) and cultured in endothelial cell growth medium (EGM) (Promocell, Heidelberg, Germany).

Growth Assay:

Article Title: Irradiation affects cellular properties and Eph receptor expression in human melanoma cells
Article Snippet: Paragraph title: Growth assay ... For growth assays, cells were precultured in T25-flasks (Greiner Bio-one), irradiated with 5 or 10 Gy and further cultured for another 6 d. Cells were plated with 1 × 105 cells per well in a 6-well plate.

Flow Cytometry:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1. .. All treatment groups were removed from culture flasks 48-h post-transfection for flow cytometry.

Microscopy:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1. .. Cells were monitored for onset of GFP using fluorescent microscopy.

Plasmid Preparation:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Paragraph title: Plasmid screening ... Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

Irradiation:

Article Title: Binding of Integrin ?6?4 to Plectin Prevents Plectin Association with F-Actin but Does Not Interfere with Intermediate Filament Binding
Article Snippet: .. The keratinocyte suspension was plated into two T25 flasks (Greiner) together with 5 × 105 irradiated 3T3 feeder cells and keratinocyte medium ( ) without EGF. ..

Article Title: Binding of Integrin ?6?4 to Plectin Prevents Plectin Association with F-Actin but Does Not Interfere with Intermediate Filament Binding
Article Snippet: .. At passage two, cells were plated in T25 flasks with irradiated 3T3 feeder cells and allowed to grow to 40–50% confluence. ..

Article Title: Irradiation affects cellular properties and Eph receptor expression in human melanoma cells
Article Snippet: .. For growth assays, cells were precultured in T25-flasks (Greiner Bio-one), irradiated with 5 or 10 Gy and further cultured for another 6 d. Cells were plated with 1 × 105 cells per well in a 6-well plate. ..

RNA Extraction:

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: .. Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction. .. All culture vessels were incubated at 37°C with 5% CO2 in a humidified incubator.

shRNA:

Article Title: Effective reduction of the interleukin-1? transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences gene expression of mediators implicated in disease pathogenesis
Article Snippet: Ten shRNA knockdown sequences and one non-targeting sequence [ ] were conceived using RNAi Block-IT Designer (Invitrogen, Carlsbad, CA) and cloned into plasmids using standard techniques. .. Chondrocytes (2.5×105 ) were seeded into T25 flasks (Greiner Bio-One, Monroe, NC) and allowed to reach ~50% confluency, at which time cultured cells were transfected using FuGENE®6 Transfection reagent (Roche Applied Science, Indianapolis, IN) in serum- and antibiotic-free DMEM according to recommendations at a reagent:DNA ratio of 3:1.

In Vitro:

Article Title: Cellular uptake, genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells
Article Snippet: An in vitro CBMN assay was carried out according to the Organization for Economic Co-operation and Development (OECD) Guideline 487 and Fenech with some modifications. .. The cells were seeded at a concentration of 2.5 × 105 cells per mL and incubated at 37 °C under CO2 atmosphere (5% CO2 in air) for 24 h in T25 flasks (Greiner Bio-one), and were treated with bare or silica-coated MNPs at a concentration range of 15–500 μg mL–1 for 4 and 24 h. The complete medium and mitomycin C (MMC; Sigma-Aldrich, USA; 0.6 μg mL–1 for 4 h and 0.3 μg mL–1 for 24 h) were used as negative and positive controls, respectively.

Concentration Assay:

Article Title: Cellular uptake, genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells
Article Snippet: .. The cells were seeded at a concentration of 2.5 × 105 cells per mL and incubated at 37 °C under CO2 atmosphere (5% CO2 in air) for 24 h in T25 flasks (Greiner Bio-one), and were treated with bare or silica-coated MNPs at a concentration range of 15–500 μg mL–1 for 4 and 24 h. The complete medium and mitomycin C (MMC; Sigma-Aldrich, USA; 0.6 μg mL–1 for 4 h and 0.3 μg mL–1 for 24 h) were used as negative and positive controls, respectively. .. At the end of the treatments, the cells were washed with complete medium, and cytochalasin B (Sigma-Aldrich, USA; final concentration of 6 μg mL–1 ) was added in the last 24 h of the culture.

Staining:

Article Title: Cellular uptake, genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells
Article Snippet: The cells were seeded at a concentration of 2.5 × 105 cells per mL and incubated at 37 °C under CO2 atmosphere (5% CO2 in air) for 24 h in T25 flasks (Greiner Bio-one), and were treated with bare or silica-coated MNPs at a concentration range of 15–500 μg mL–1 for 4 and 24 h. The complete medium and mitomycin C (MMC; Sigma-Aldrich, USA; 0.6 μg mL–1 for 4 h and 0.3 μg mL–1 for 24 h) were used as negative and positive controls, respectively. .. The cells were then fixed with methanol–acetic acid (3 : 1 v/v), dropped onto cold slides, air-dried and stained with Giemsa–May Grünwald solution (Merck).

Article Title: Potency Biomarker Signature Genes from Multiparametric Osteogenesis Assays: Will cGMP Human Bone Marrow Mesenchymal Stromal Cells Make Bone?
Article Snippet: .. Cells were plated at a seeding density of 104 /cm2 in 24-well multiwell plates (Greiner Bio-one) for Alizarin Red staining and in T25 flasks (Greiner Bio-one) for RNA extraction. .. All culture vessels were incubated at 37°C with 5% CO2 in a humidified incubator.

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  • 90
    Greiner Bio t25 flask
    Analysis of colony distributions after 12 Gy at CSU. ( A ) Uneven colony distribution with increased survival near the wall of the flasks for 4 MeV compared to 18 MeV electron irradiation. 100,000 cells were plated to <t>T25</t> flasks, T75 flask and glass dishes. ( B ) Increased colony survival near the edge of the 6 cm plastic dish after 25 J/m 2 of UV-C exposure. ( C ) Quantitative analysis of colony distribution using 4, 9, and 18 MeV. 100,000 cells were plated. Error bars indicate the standard error of the means for three independent experiments. Note the increased number of surviving colonies near the edges of the flask at 4 MeV.
    T25 Flask, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 90/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t25 flask/product/Greiner Bio
    Average 90 stars, based on 28 article reviews
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    Greiner Bio t25 culture flasks
    Alteration in protein levels and phosphorylation on short term AZD8055 treatment Cells at CPD 36 or CPD 73 were seeded into <t>T25</t> flasks and grown in medium containing AZD8055 or DMSO for one week. After harvesting, cells were pelleted and lysed in RIPA with protease and phosphatase inhibitors, separated on SDS-PAGE then immunoblotted for the indicated proteins. ( A ) Components of the mTOR signaling pathway, ( B ) Cell cycle proteins. The panels on the right show longer exposure images of the same panels on the left, to highlight bands that are faint in the shorter exposure images.
    T25 Culture Flasks, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t25 culture flasks/product/Greiner Bio
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t25 culture flasks - by Bioz Stars, 2020-04
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    85
    Greiner Bio t25 tissue flasks
    (a)–(c) Photomicrographs of NPI after culture for 1 (a), 6 (b), and 10 (c) days on <t>T25</t> tissue flasks for adherent cell growth. (d) Light field (left) and fluorescence (right) photomicrographs of Ba-AG microcapsules containing SC. Fluorescence micrographs were obtained after staining with EB+FDA to assess SC viability. (e) Insulin secretory patterns of control NPI cell monolayers alone after 14 days (open bars) or NPI cell monolayers, cocultivated with microencapsulated SC for 7 (gray bars), 14 (hatched bars), and 21 days (filled bars) of culture. During static incubation, the cells were treated with the indicated concentrations of glucose. Data represent the average of 3 independent experiments; each insulin determination was performed in triplicate ±SD.
    T25 Tissue Flasks, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t25 tissue flasks/product/Greiner Bio
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t25 tissue flasks - by Bioz Stars, 2020-04
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    Image Search Results


    Analysis of colony distributions after 12 Gy at CSU. ( A ) Uneven colony distribution with increased survival near the wall of the flasks for 4 MeV compared to 18 MeV electron irradiation. 100,000 cells were plated to T25 flasks, T75 flask and glass dishes. ( B ) Increased colony survival near the edge of the 6 cm plastic dish after 25 J/m 2 of UV-C exposure. ( C ) Quantitative analysis of colony distribution using 4, 9, and 18 MeV. 100,000 cells were plated. Error bars indicate the standard error of the means for three independent experiments. Note the increased number of surviving colonies near the edges of the flask at 4 MeV.

    Journal: Scientific Reports

    Article Title: Electron Scattering in Conventional Cell Flask Experiments and Dose Distribution Dependency

    doi: 10.1038/s41598-019-57029-y

    Figure Lengend Snippet: Analysis of colony distributions after 12 Gy at CSU. ( A ) Uneven colony distribution with increased survival near the wall of the flasks for 4 MeV compared to 18 MeV electron irradiation. 100,000 cells were plated to T25 flasks, T75 flask and glass dishes. ( B ) Increased colony survival near the edge of the 6 cm plastic dish after 25 J/m 2 of UV-C exposure. ( C ) Quantitative analysis of colony distribution using 4, 9, and 18 MeV. 100,000 cells were plated. Error bars indicate the standard error of the means for three independent experiments. Note the increased number of surviving colonies near the edges of the flask at 4 MeV.

    Article Snippet: This was carried out using T25 flask, the T75 flask (Greiner Bio-One), P-60 6 cm dish (Greiner Bio-One), 6 wells plate (Greiner Bio-One), and Pyrex 60 mm glass dish (Pyrex, Corning, NY) in order to remove the flask size and shape influence (Fig. ).

    Techniques: Irradiation

    Detection of Venus-fluorescence and A . t .-sequences in recipient cells after passaging. (a) 2x10 5 hMSC were seeded into T25, incubated overnight to reach adherence (d0) and fed with EV derived from A . t .-hMSC cultures for 2 weeks. (b) Venus-positive cells were detected in 2 of 4 flasks (d14). One flask with 7 positive cells was passaged into 4xT25 flasks. (c) 7 days later (d21), one flask contained 10 Venus-positive cells. This culture was expanded again into 4xT25. (d) Venus-positive cells at d28 were evident in 2 flasks out of 4 with 13 cells in one flask and 1 cell in the second flask. Exemplarily, one positive MSC spot with corresponding phase-contrast for each time point is shown. Magnification x 100. (e, f) DNA of the flask with 13 Venus-positive cells was pretested in nested SYBR Green-based qPCR. Out of 10 primary reaction tubes, 4 were positive in the nested qPCR tested in 8 replicates (tubes 2, 4, 7 and 8; not shown) and were retested in TaqMan-based qPCR (e) and ddPCR (f). Shown are the results for positive control (pc, 10 copies/PCR reaction; 4 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), negative control (nc, untransduced hMSC; 8 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), and tube 2 and 4 (16 replicates in TaqMan-based qPCR and ddPCR).

    Journal: PLoS ONE

    Article Title: Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles

    doi: 10.1371/journal.pone.0163665

    Figure Lengend Snippet: Detection of Venus-fluorescence and A . t .-sequences in recipient cells after passaging. (a) 2x10 5 hMSC were seeded into T25, incubated overnight to reach adherence (d0) and fed with EV derived from A . t .-hMSC cultures for 2 weeks. (b) Venus-positive cells were detected in 2 of 4 flasks (d14). One flask with 7 positive cells was passaged into 4xT25 flasks. (c) 7 days later (d21), one flask contained 10 Venus-positive cells. This culture was expanded again into 4xT25. (d) Venus-positive cells at d28 were evident in 2 flasks out of 4 with 13 cells in one flask and 1 cell in the second flask. Exemplarily, one positive MSC spot with corresponding phase-contrast for each time point is shown. Magnification x 100. (e, f) DNA of the flask with 13 Venus-positive cells was pretested in nested SYBR Green-based qPCR. Out of 10 primary reaction tubes, 4 were positive in the nested qPCR tested in 8 replicates (tubes 2, 4, 7 and 8; not shown) and were retested in TaqMan-based qPCR (e) and ddPCR (f). Shown are the results for positive control (pc, 10 copies/PCR reaction; 4 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), negative control (nc, untransduced hMSC; 8 replicates in TaqMan-based qPCR and 2 replicates in ddPCR), and tube 2 and 4 (16 replicates in TaqMan-based qPCR and ddPCR).

    Article Snippet: EV-transfer to recipient cells Recipient human BM-hMSC (2.5-5x104 ) unrelated to hMSC used for A .t .-transduction were seeded into T25-flasks (Greiner).

    Techniques: Fluorescence, Passaging, Incubation, Derivative Assay, SYBR Green Assay, Real-time Polymerase Chain Reaction, Positive Control, Polymerase Chain Reaction, Negative Control

    Alteration in protein levels and phosphorylation on short term AZD8055 treatment Cells at CPD 36 or CPD 73 were seeded into T25 flasks and grown in medium containing AZD8055 or DMSO for one week. After harvesting, cells were pelleted and lysed in RIPA with protease and phosphatase inhibitors, separated on SDS-PAGE then immunoblotted for the indicated proteins. ( A ) Components of the mTOR signaling pathway, ( B ) Cell cycle proteins. The panels on the right show longer exposure images of the same panels on the left, to highlight bands that are faint in the shorter exposure images.

    Journal: Aging (Albany NY)

    Article Title: Reversal of phenotypes of cellular senescence by pan-mTOR inhibition

    doi:

    Figure Lengend Snippet: Alteration in protein levels and phosphorylation on short term AZD8055 treatment Cells at CPD 36 or CPD 73 were seeded into T25 flasks and grown in medium containing AZD8055 or DMSO for one week. After harvesting, cells were pelleted and lysed in RIPA with protease and phosphatase inhibitors, separated on SDS-PAGE then immunoblotted for the indicated proteins. ( A ) Components of the mTOR signaling pathway, ( B ) Cell cycle proteins. The panels on the right show longer exposure images of the same panels on the left, to highlight bands that are faint in the shorter exposure images.

    Article Snippet: Immunoblotting Cells were seeded at 2×105 in T25 culture flasks (Greiner) and harvested by trypsinization as above, then pelleted by gentle centrifugation, washed in PBS and re-pelleted, then lysed in 30 μl RIPA buffer containing 1:100 HaltTM protease and phosphatase inhibitors (Thermo Scientific).

    Techniques: SDS Page

    (a)–(c) Photomicrographs of NPI after culture for 1 (a), 6 (b), and 10 (c) days on T25 tissue flasks for adherent cell growth. (d) Light field (left) and fluorescence (right) photomicrographs of Ba-AG microcapsules containing SC. Fluorescence micrographs were obtained after staining with EB+FDA to assess SC viability. (e) Insulin secretory patterns of control NPI cell monolayers alone after 14 days (open bars) or NPI cell monolayers, cocultivated with microencapsulated SC for 7 (gray bars), 14 (hatched bars), and 21 days (filled bars) of culture. During static incubation, the cells were treated with the indicated concentrations of glucose. Data represent the average of 3 independent experiments; each insulin determination was performed in triplicate ±SD.

    Journal: Stem Cells International

    Article Title: Acceleration of Functional Maturation and Differentiation of Neonatal Porcine Islet Cell Monolayers Shortly In Vitro Cocultured with Microencapsulated Sertoli Cells

    doi: 10.4061/2010/587213

    Figure Lengend Snippet: (a)–(c) Photomicrographs of NPI after culture for 1 (a), 6 (b), and 10 (c) days on T25 tissue flasks for adherent cell growth. (d) Light field (left) and fluorescence (right) photomicrographs of Ba-AG microcapsules containing SC. Fluorescence micrographs were obtained after staining with EB+FDA to assess SC viability. (e) Insulin secretory patterns of control NPI cell monolayers alone after 14 days (open bars) or NPI cell monolayers, cocultivated with microencapsulated SC for 7 (gray bars), 14 (hatched bars), and 21 days (filled bars) of culture. During static incubation, the cells were treated with the indicated concentrations of glucose. Data represent the average of 3 independent experiments; each insulin determination was performed in triplicate ±SD.

    Article Snippet: NPI were than replated in Click's medium (Sigma Chemical Co) supplemented with 10% fetal bovine serum (EuroClone, Wetherby, UK), 0.5% bovine serum albumin, fraction V (BSA) (Sigma Chemical Co), 10 mM nicotinamide (Sigma Chemical Co), 2 mM L-glutamine (Sigma Chemical Co), and penicillin +0.1 mg/mL streptomycin (Sigma Chemical Co) using T25 tissue flasks for adherent cell growth (Greiner Bio-one, Frickenhausen, Germany), at a concentration of 30 NPI/flask.

    Techniques: Fluorescence, Staining, Incubation