non-essential amino-acid Search Results


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  • 99
    Thermo Fisher non essential amino acids
    (A) Schema of genome editing for ACLY . A GFP-tagged sgRNA/Cas9 plasmid targeting ACLY was transfected into HT1080 cells. Following cell sorting by flow cytometry for GFP positive transfected cells, cells were recovered in either standard medium (no added acetate) or 2 or 20 mM sodium acetate-supplemented media. Recovered cell clones were genotyped to identify genome editing for ACLY as in “B”. (B) Schema of genotyping for ACLY genome-edited cell clones and representative genotyping results (top left). The sgRNA-targeted and Cas9 cleavage site in ACLY exon 6, which contains a single Xho I site overlapped with a Cas9 cleavage site, was PCR amplified by flanked primers (blue). Xho I digestion of the PCR product (385 bp) provided 238 bp and 147 bp fragments when the Xho I site was intact (top right). Xho I resistance was indicative of insertion or deletion (in/del) by CRISPR-mediated <t>non-homologous</t> end joining (NHEJ). Numbers of clones which exhibited complete (or partial) Xho I resistance and numbers of clones screened were shown for each culture media condition (bottom). (C) The sequence of exon 6 in wildtype ACLY and the edited sequences in ASA-KO14 and KO17 cells. The sgRNA target site (red), PAM motif (green), Cas9 cut site (arrow head), Xho I site, and detected in/del were indicated. We detected only a single pattern of insertion in ASA-KO14 cells. Note that one allele of ASA-KO17 possesses an indel which causes a frame shift mutation replacing <t>amino</t> <t>acids</t> “T-Y-I-E” with “Q”. This might explain the detected ACLY peptides in the TMT proteomics using ASA-KO17 cells (Table S3).
    Non Essential Amino Acids, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 27022 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5847 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher non essential amino acid
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acid, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1421 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 1x non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    1x Non Essential Amino Acids, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 737 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 961 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mediatech non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Mediatech, used in various techniques. Bioz Stars score: 93/100, based on 458 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biochrom non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Biochrom, used in various techniques. Bioz Stars score: 93/100, based on 514 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 247 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore non essential amino acid solution
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acid Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 367 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cellgro non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Cellgro, used in various techniques. Bioz Stars score: 92/100, based on 337 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biological Industries Inc non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by Biological Industries Inc, used in various techniques. Bioz Stars score: 92/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PAA Laboratories non essential amino acids
    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% <t>non-essential</t> <t>amino</t> <t>acids,</t> 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p
    Non Essential Amino Acids, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 94/100, based on 292 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    (A) Schema of genome editing for ACLY . A GFP-tagged sgRNA/Cas9 plasmid targeting ACLY was transfected into HT1080 cells. Following cell sorting by flow cytometry for GFP positive transfected cells, cells were recovered in either standard medium (no added acetate) or 2 or 20 mM sodium acetate-supplemented media. Recovered cell clones were genotyped to identify genome editing for ACLY as in “B”. (B) Schema of genotyping for ACLY genome-edited cell clones and representative genotyping results (top left). The sgRNA-targeted and Cas9 cleavage site in ACLY exon 6, which contains a single Xho I site overlapped with a Cas9 cleavage site, was PCR amplified by flanked primers (blue). Xho I digestion of the PCR product (385 bp) provided 238 bp and 147 bp fragments when the Xho I site was intact (top right). Xho I resistance was indicative of insertion or deletion (in/del) by CRISPR-mediated non-homologous end joining (NHEJ). Numbers of clones which exhibited complete (or partial) Xho I resistance and numbers of clones screened were shown for each culture media condition (bottom). (C) The sequence of exon 6 in wildtype ACLY and the edited sequences in ASA-KO14 and KO17 cells. The sgRNA target site (red), PAM motif (green), Cas9 cut site (arrow head), Xho I site, and detected in/del were indicated. We detected only a single pattern of insertion in ASA-KO14 cells. Note that one allele of ASA-KO17 possesses an indel which causes a frame shift mutation replacing amino acids “T-Y-I-E” with “Q”. This might explain the detected ACLY peptides in the TMT proteomics using ASA-KO17 cells (Table S3).

    Journal: bioRxiv

    Article Title: Acetylation-mediated phase control of the nucleolus regulates cellular acetyl-CoA responses

    doi: 10.1101/2020.01.24.918706

    Figure Lengend Snippet: (A) Schema of genome editing for ACLY . A GFP-tagged sgRNA/Cas9 plasmid targeting ACLY was transfected into HT1080 cells. Following cell sorting by flow cytometry for GFP positive transfected cells, cells were recovered in either standard medium (no added acetate) or 2 or 20 mM sodium acetate-supplemented media. Recovered cell clones were genotyped to identify genome editing for ACLY as in “B”. (B) Schema of genotyping for ACLY genome-edited cell clones and representative genotyping results (top left). The sgRNA-targeted and Cas9 cleavage site in ACLY exon 6, which contains a single Xho I site overlapped with a Cas9 cleavage site, was PCR amplified by flanked primers (blue). Xho I digestion of the PCR product (385 bp) provided 238 bp and 147 bp fragments when the Xho I site was intact (top right). Xho I resistance was indicative of insertion or deletion (in/del) by CRISPR-mediated non-homologous end joining (NHEJ). Numbers of clones which exhibited complete (or partial) Xho I resistance and numbers of clones screened were shown for each culture media condition (bottom). (C) The sequence of exon 6 in wildtype ACLY and the edited sequences in ASA-KO14 and KO17 cells. The sgRNA target site (red), PAM motif (green), Cas9 cut site (arrow head), Xho I site, and detected in/del were indicated. We detected only a single pattern of insertion in ASA-KO14 cells. Note that one allele of ASA-KO17 possesses an indel which causes a frame shift mutation replacing amino acids “T-Y-I-E” with “Q”. This might explain the detected ACLY peptides in the TMT proteomics using ASA-KO17 cells (Table S3).

    Article Snippet: 293T cells (Lenti-X, Takara, San Francisco, CA) and HCT116 cells ( )were cultured in the same condition as HT1080 cells but in Dulbeccos’s Modified Eagle Medium (Gibco) supplemented with 10% FBS, 1 mM Sodium pyruvate (Gibco), non-essential amino acids (Gibco) and GlutaMAX (Gibco).

    Techniques: Plasmid Preparation, Transfection, FACS, Flow Cytometry, Clone Assay, Polymerase Chain Reaction, Amplification, CRISPR, Non-Homologous End Joining, Sequencing, Mutagenesis

    Saporin-encapsulated exosomes enhances cytotoxicity by EGF stimulation and macropinocytosis induction. ( a ) Cell viability of A431 cells treated with CD63-GFP-exosomes (20 μg/ml) in serum-free cell culture medium with or without co-treatment of EGF (100 or 500 nM) for 24 h at 37 °C, analysed using a WST-1 assay. ( b ) Cytotoxicity of saporin-encapsulated exosomes (4 μg/ml) or saporin (7 μg/ml) with or without co-treatment of EGF (500 nM). A431 cells were treated with each test sample in 10% FBS containing cell culture medium for 48 h at 37 °C, prior to a WST-1 assay. The data are the averages (±SD) of three experiments. ** p

    Journal: Scientific Reports

    Article Title: Active macropinocytosis induction by stimulation of epidermal growth factor receptor and oncogenic Ras expression potentiates cellular uptake efficacy of exosomes

    doi: 10.1038/srep10300

    Figure Lengend Snippet: Saporin-encapsulated exosomes enhances cytotoxicity by EGF stimulation and macropinocytosis induction. ( a ) Cell viability of A431 cells treated with CD63-GFP-exosomes (20 μg/ml) in serum-free cell culture medium with or without co-treatment of EGF (100 or 500 nM) for 24 h at 37 °C, analysed using a WST-1 assay. ( b ) Cytotoxicity of saporin-encapsulated exosomes (4 μg/ml) or saporin (7 μg/ml) with or without co-treatment of EGF (500 nM). A431 cells were treated with each test sample in 10% FBS containing cell culture medium for 48 h at 37 °C, prior to a WST-1 assay. The data are the averages (±SD) of three experiments. ** p

    Article Snippet: The cells were cultured in minimum essential medium α (α-MEM; Gibco, Life Technologies Corporation, Grand Island, NY, USA) containing 10% heat-inactivated foetal bovine serum (FBS; Gibco, Life Technologies Corporation) (HeLa cells), minimum essential medium (MEM) (Gibco, Life Technologies Corporation) containing 10% heat-inactivated FBS (Gibco, Life Technologies Corporation) (A431 cells), Eagle’s minimum essential medium (EMEM) (Wako, Osaka, Japan) containing 10% FBS (HyClone Laboratories, South Logan, UT, USA), 0.1 mM MEM non-essential amino acids (Gibco), penicillin-streptomycin (50 units/ml and 50 μg/ml) (Sigma-Aldrich, St. Louis, MO, USA) (MIA PaCa-2 cells), and RPMI1640 (Gibco, Life Technologies Corporation) containing 10% FBS (HyClone), and penicillin-streptomycin (50 units/ml and 50 μg/ml) (Sigma-Aldrich) (BxPC-3 cells).

    Techniques: Cell Culture, WST-1 Assay

    Effective cytotoxicity of saporin-EGF-encapsulated exosomes. ( a ) Microscopic observation of A431 cells treated with saporin- or saporin-EGF-encapsulated exosomes (0.4, 4, 20 μg/ml) for 72 h at 37 °C in 10% FBS-containing cell culture medium. ( b ) Cell viability of A431 cells treated with saporin- (blue line) or saporin-EGF- (red line) encapsulated exosomes (0.4, 4, 20 μg/ml) in the same experimental condition of ( a ) analysed using a WST-1 assay. The data represent the average (±SD) of four experiments. *** p

    Journal: Scientific Reports

    Article Title: Active macropinocytosis induction by stimulation of epidermal growth factor receptor and oncogenic Ras expression potentiates cellular uptake efficacy of exosomes

    doi: 10.1038/srep10300

    Figure Lengend Snippet: Effective cytotoxicity of saporin-EGF-encapsulated exosomes. ( a ) Microscopic observation of A431 cells treated with saporin- or saporin-EGF-encapsulated exosomes (0.4, 4, 20 μg/ml) for 72 h at 37 °C in 10% FBS-containing cell culture medium. ( b ) Cell viability of A431 cells treated with saporin- (blue line) or saporin-EGF- (red line) encapsulated exosomes (0.4, 4, 20 μg/ml) in the same experimental condition of ( a ) analysed using a WST-1 assay. The data represent the average (±SD) of four experiments. *** p

    Article Snippet: The cells were cultured in minimum essential medium α (α-MEM; Gibco, Life Technologies Corporation, Grand Island, NY, USA) containing 10% heat-inactivated foetal bovine serum (FBS; Gibco, Life Technologies Corporation) (HeLa cells), minimum essential medium (MEM) (Gibco, Life Technologies Corporation) containing 10% heat-inactivated FBS (Gibco, Life Technologies Corporation) (A431 cells), Eagle’s minimum essential medium (EMEM) (Wako, Osaka, Japan) containing 10% FBS (HyClone Laboratories, South Logan, UT, USA), 0.1 mM MEM non-essential amino acids (Gibco), penicillin-streptomycin (50 units/ml and 50 μg/ml) (Sigma-Aldrich, St. Louis, MO, USA) (MIA PaCa-2 cells), and RPMI1640 (Gibco, Life Technologies Corporation) containing 10% FBS (HyClone), and penicillin-streptomycin (50 units/ml and 50 μg/ml) (Sigma-Aldrich) (BxPC-3 cells).

    Techniques: Cell Culture, WST-1 Assay

    TA-LF complexes promotes delivery of encapsulated therapeutic drugs to LC cells. MTS assay of drug-encapsulated TA-LF complexes against ( a ) A540 and ( b ) H1299 LC cells. Cells (5 × 103) were seeded in a 96-well plate and left overnight for cell attachment to the plate , the cells were treated with indicated concentrations of gemcitabine, carboplatin, and irinotecan and their respective drug-encapsulated TA-LF complexes for 48 h. Cell viability was determined using MTS assay. TA at the concentrations used to make TA-LF complexes were used as controls. The data were presented in the form of line graphs as percent viable cells compared to untreated cells in medium. Data presented as mean ± SEM (each treatment, n = 6). Cytotoxicity of TA-LF-drug formulations were significant compared to free drugs (* p

    Journal: Pharmaceutics

    Article Title: Tannic Acid-Lung Fluid Assemblies Promote Interaction and Delivery of Drugs to Lung Cancer Cells

    doi: 10.3390/pharmaceutics10030111

    Figure Lengend Snippet: TA-LF complexes promotes delivery of encapsulated therapeutic drugs to LC cells. MTS assay of drug-encapsulated TA-LF complexes against ( a ) A540 and ( b ) H1299 LC cells. Cells (5 × 103) were seeded in a 96-well plate and left overnight for cell attachment to the plate , the cells were treated with indicated concentrations of gemcitabine, carboplatin, and irinotecan and their respective drug-encapsulated TA-LF complexes for 48 h. Cell viability was determined using MTS assay. TA at the concentrations used to make TA-LF complexes were used as controls. The data were presented in the form of line graphs as percent viable cells compared to untreated cells in medium. Data presented as mean ± SEM (each treatment, n = 6). Cytotoxicity of TA-LF-drug formulations were significant compared to free drugs (* p

    Article Snippet: The cancer cell lines cultured under sterile condition in Dulbecco’s Modified Eagle’s medium (DMEM for A549) and Roswell Park Memorial Institute medium (RPMI for H1299) supplemented with 4.5 g/L of glucose, 10 nM of nonessential amino acids (# 11140076, Gibco, Thermo Fisher Scientific, Grand Island, NY, USA), 100 mM of sodium pyruvate (#11360070, Gibco), 1× antibiotic/antimycotic (#15240062, Gibco), and 10% heat-inactivated FBS (#10438026 Thermo Fisher).

    Techniques: MTS Assay, Cell Attachment Assay

    Horse milk is toxic for C. coli . Milk susceptibility of A) C. jejuni or B) C. coli to horse, cow, or goat milk determined by incubating overnight at 37 °C in a microaerophilic environment 1.0 × 10 9 CFU Campylobacter bacteria in 1 ml DMEM medium + 10% FBS + 1× NEAA–milk (1:1) ratio. Scatter plot shows the mean and standard error of the mean of three independent experiments

    Journal: European Journal of Microbiology & Immunology

    Article Title: Milk Modulates Campylobacter Invasion into Caco-2 Intestinal Epithelial Cells

    doi: 10.1556/1886.2015.00019

    Figure Lengend Snippet: Horse milk is toxic for C. coli . Milk susceptibility of A) C. jejuni or B) C. coli to horse, cow, or goat milk determined by incubating overnight at 37 °C in a microaerophilic environment 1.0 × 10 9 CFU Campylobacter bacteria in 1 ml DMEM medium + 10% FBS + 1× NEAA–milk (1:1) ratio. Scatter plot shows the mean and standard error of the mean of three independent experiments

    Article Snippet: After recovery, cells were harvested in Dulbecco’s modified Eagle medium (DMEM) (Life Technology, Breda, The Netherlands) containing 10% fetal bovine serum (FBS) (Life Technology, Breda, The Netherlands) and 1× non-essential amino acids (NEAA) (Life Technology, Breda, The Netherlands), and densities were adjusted according to the optical density at an OD600 nm where 1 OD equals 2.5 × 1009 CFU/ml.

    Techniques:

    Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% non-essential amino acids, 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p

    Journal: Biomolecules

    Article Title: Expression of SCD and FADS2 Is Lower in the Necrotic Core and Growing Tumor Area than in the Peritumoral Area of Glioblastoma Multiforme

    doi: 10.3390/biom10050727

    Figure Lengend Snippet: Effect of the necrotic condition or nutritional deficiency on the mRNA expression of FADS1 ( a , b ), FADS2 ( c , d ), and FADS3 ( e , f ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in EMEM medium with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, 1% non-essential amino acids, 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with PBS solution. Next, cells were cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, while starved cells were grown in medium with low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p

    Article Snippet: Cell Culture and Treatment Human brain cells (glioblastoma astrocytoma, U-87 MG cell line) from the European Collection of Authenticated Cell Cultures (ECACC) were cultured in eagle’s minimum essential medium (EMEM), (Sigma-Aldrich, Poznań, Poland) supplemented with 10% (v /v ) heat-inactivated fetal bovine serum (FBS; Gibco Limited, Poznań Poland), 2 mM l -glutamine, 1 mM sodium pyruvate (Sigma-Aldrich, Poznań, Poland), 1% non-essential amino acids (Sigma-Aldrich, Poznań, Poland), 100 U/mL penicillin (Gibco Limited, Poznań, Poland), and 100 µg/mL streptomycin (Gibco Limited, Poznań, Poland), at 37 °C in a humidified atmosphere of 95% air and 5% CO2 .

    Techniques: Expressing, Cell Culture, Incubation, Concentration Assay, Quantitative RT-PCR

    The effect of necrotic condition or nutritional deficiency on the mRNA expression of SCD ( a , b ) and SCD5 ( c , d ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in eagle’s minimum essential medium (EMEM) with 10% fetal bovine serum (FBS), 2 mM l -glutamine, 1 mM sodium pyruvate, 1% non essential amino acids, 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with phosphate-buffered saline (PBS) solution and cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, starved cells were grown in medium with a low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p

    Journal: Biomolecules

    Article Title: Expression of SCD and FADS2 Is Lower in the Necrotic Core and Growing Tumor Area than in the Peritumoral Area of Glioblastoma Multiforme

    doi: 10.3390/biom10050727

    Figure Lengend Snippet: The effect of necrotic condition or nutritional deficiency on the mRNA expression of SCD ( a , b ) and SCD5 ( c , d ). Human brain (glioblastoma astrocytoma) U-87 MG cell line cultured in eagle’s minimum essential medium (EMEM) with 10% fetal bovine serum (FBS), 2 mM l -glutamine, 1 mM sodium pyruvate, 1% non essential amino acids, 100 U/mL penicillin, and 100 µg/mL streptomycin. U-87 MG cells were seeded in 6-well plates. After 72 h incubation, cells were washed three times with phosphate-buffered saline (PBS) solution and cultured for 24 h under three different conditions (control, nutrient deficiency, and necrotic). Control cells were suspended in full medium, starved cells were grown in medium with a low concentration of l -glutamine (0.2 mM) and without sodium pyruvate. For the induction of necrotic conditions, cells were incubated in medium supplemented with 200 µM H 2 O 2 . After 24 h of incubation, the cells were trypsinized and analyzed using qRT-PCR. **—statistically significant difference in the expression of a given desaturase between the tumor area according to the Wilcoxon signed-rank test ( p

    Article Snippet: Cell Culture and Treatment Human brain cells (glioblastoma astrocytoma, U-87 MG cell line) from the European Collection of Authenticated Cell Cultures (ECACC) were cultured in eagle’s minimum essential medium (EMEM), (Sigma-Aldrich, Poznań, Poland) supplemented with 10% (v /v ) heat-inactivated fetal bovine serum (FBS; Gibco Limited, Poznań Poland), 2 mM l -glutamine, 1 mM sodium pyruvate (Sigma-Aldrich, Poznań, Poland), 1% non-essential amino acids (Sigma-Aldrich, Poznań, Poland), 100 U/mL penicillin (Gibco Limited, Poznań, Poland), and 100 µg/mL streptomycin (Gibco Limited, Poznań, Poland), at 37 °C in a humidified atmosphere of 95% air and 5% CO2 .

    Techniques: Expressing, Cell Culture, Incubation, Concentration Assay, Quantitative RT-PCR

    Initial provision of serum, bFGF or SCF is not necessary for conversion of mouse PGCs into EG cells in presence of 2i-LIF. ( A ) Schematic of derivation protocol to test requirement for FCS and bFGF. Primordial germ cells (PGCs) are plated either directly into 2i-LIF only or into FCS-LIF plus bFGF. ( B ) Quantitation of EG cell colony formation following derivation directly into 2i-LIF or transfer to 2i-LIF after the first medium change. ( C ) Schematic of derivation protocol to test requirement for stem cell factor (SCF). PGCs were plated directly into 2i-LIF either onto Sl 4 -m220 feeders, Sl/Sl 4 feeders or Sl/Sl 4 feeders with the addition of soluble SCF. ( D ) Quantitation of EG cell colony formation following derivation directly into 2i-LIF in the presence of membrane-bound SCF, no SCF or soluble SCF.

    Journal: Development (Cambridge, England)

    Article Title: Embryonic germ cells from mice and rats exhibit properties consistent with a generic pluripotent ground state

    doi: 10.1242/dev.050427

    Figure Lengend Snippet: Initial provision of serum, bFGF or SCF is not necessary for conversion of mouse PGCs into EG cells in presence of 2i-LIF. ( A ) Schematic of derivation protocol to test requirement for FCS and bFGF. Primordial germ cells (PGCs) are plated either directly into 2i-LIF only or into FCS-LIF plus bFGF. ( B ) Quantitation of EG cell colony formation following derivation directly into 2i-LIF or transfer to 2i-LIF after the first medium change. ( C ) Schematic of derivation protocol to test requirement for stem cell factor (SCF). PGCs were plated directly into 2i-LIF either onto Sl 4 -m220 feeders, Sl/Sl 4 feeders or Sl/Sl 4 feeders with the addition of soluble SCF. ( D ) Quantitation of EG cell colony formation following derivation directly into 2i-LIF in the presence of membrane-bound SCF, no SCF or soluble SCF.

    Article Snippet: FCS-LIF conditions comprise DMEM-F12 medium supplemented with 15% FCS, 0.1% MEM non-essential amino acids, 4 mM glutamate, 2 mM sodium pyruvate, 0.1 mM 2-mecaptanethanol and recombinant mouse LIF (1200 U/ml; ESGRO, Chemicon) or human recombinant LIF generated in-house by transient transfection of Cos7 cells with plasmid encoding human LIF.

    Techniques: Quantitation Assay

    Mouse EG cells can be maintained and derived in 2i-LIF. ( A ) Phase contrast and fluorescence images of Oct4-ΔPE-GFP EG cells. ( B ) Phase contrast and fluorescence images of Oct4-ΔPE-GFP EG cells cultured in: FCS-LIF on a MEF feeder layer; FCS-LIF after one passage without feeders; 2i-LIF without feeders. ( C ) Flow cytometry analysis of Oct4-ΔPE-GFP EG cells cultured in 2i-LIF compared with FCS-LIF on feeders. Gates were set to eliminate feeders and cell debris. ( D ) Colony-forming assay on Oct4-ΔPE-GFP EG cells. Colonies were scored for alkaline phosphatase (AP) and designated positive, negative or mixed. Data are means of two biological replicates. ( E ) Schematic of EG cell derivation protocol. On day 3, medium was changed to either FCS-LIF or 2i-LIF. ( F ) Comparison of GFP-positive EG cell colony formation following addition of 2i-LIF on day 3 compared with FCS-LIF. ( G ) High-contribution coat colour chimaeras generated with agouti EG cells derived in 2i-LIF injected into C57BL/6 blastocysts. ( H ) Chimaera, C57BL/6 mate, and mixed litter of agouti and black pups. Scale bars: 100 μm in A; 25 μm in B.

    Journal: Development (Cambridge, England)

    Article Title: Embryonic germ cells from mice and rats exhibit properties consistent with a generic pluripotent ground state

    doi: 10.1242/dev.050427

    Figure Lengend Snippet: Mouse EG cells can be maintained and derived in 2i-LIF. ( A ) Phase contrast and fluorescence images of Oct4-ΔPE-GFP EG cells. ( B ) Phase contrast and fluorescence images of Oct4-ΔPE-GFP EG cells cultured in: FCS-LIF on a MEF feeder layer; FCS-LIF after one passage without feeders; 2i-LIF without feeders. ( C ) Flow cytometry analysis of Oct4-ΔPE-GFP EG cells cultured in 2i-LIF compared with FCS-LIF on feeders. Gates were set to eliminate feeders and cell debris. ( D ) Colony-forming assay on Oct4-ΔPE-GFP EG cells. Colonies were scored for alkaline phosphatase (AP) and designated positive, negative or mixed. Data are means of two biological replicates. ( E ) Schematic of EG cell derivation protocol. On day 3, medium was changed to either FCS-LIF or 2i-LIF. ( F ) Comparison of GFP-positive EG cell colony formation following addition of 2i-LIF on day 3 compared with FCS-LIF. ( G ) High-contribution coat colour chimaeras generated with agouti EG cells derived in 2i-LIF injected into C57BL/6 blastocysts. ( H ) Chimaera, C57BL/6 mate, and mixed litter of agouti and black pups. Scale bars: 100 μm in A; 25 μm in B.

    Article Snippet: FCS-LIF conditions comprise DMEM-F12 medium supplemented with 15% FCS, 0.1% MEM non-essential amino acids, 4 mM glutamate, 2 mM sodium pyruvate, 0.1 mM 2-mecaptanethanol and recombinant mouse LIF (1200 U/ml; ESGRO, Chemicon) or human recombinant LIF generated in-house by transient transfection of Cos7 cells with plasmid encoding human LIF.

    Techniques: Derivative Assay, Fluorescence, Cell Culture, Flow Cytometry, Cytometry, Generated, Injection

    The morphology and RT-PCR analyses of the expression of insulin producing cells-related genes in P-ADSCs during the induced differentiation process. The step 1 decrease the FBS concentration from 10% to 2% and supplemented wtih NEAA and β-mercaptoethanol, then step 2 supplement with activin A, nicotinamide and bFGF, EGF, finally step 3 activin A, nicotinamide, and exendin 4 was supplied. a The changes in cell morphology could be observed from day 2 to day 16 after induction. b Expression analysis of islet markers using RT-PCR in pADSC after β-cell differentiation with indicated protocols. Scale bar 100 μm for micrograph

    Journal: Cytotechnology

    Article Title: Investigating the mincing method for isolation of adipose-derived stem cells from pregnant women fat

    doi: 10.1007/s10616-017-0162-8

    Figure Lengend Snippet: The morphology and RT-PCR analyses of the expression of insulin producing cells-related genes in P-ADSCs during the induced differentiation process. The step 1 decrease the FBS concentration from 10% to 2% and supplemented wtih NEAA and β-mercaptoethanol, then step 2 supplement with activin A, nicotinamide and bFGF, EGF, finally step 3 activin A, nicotinamide, and exendin 4 was supplied. a The changes in cell morphology could be observed from day 2 to day 16 after induction. b Expression analysis of islet markers using RT-PCR in pADSC after β-cell differentiation with indicated protocols. Scale bar 100 μm for micrograph

    Article Snippet: The first step of the method was seeding P-ADSCs into a 100 mm dish (1 x106 cells/dish) containing 2% FBS/DMEM (high glucose) supplemented with 1% non-essential amino acids (NEAA) and 0.5 mM β-mercaptoethanol (Sigma) for 2 days.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Concentration Assay, Cell Differentiation