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  • 99
    Thermo Fisher dmem f12
    The effect of solution added TGF-β in <t>DMEM/F12/10%</t> FBS media + DMSO on CSPG production. CSPG staining per cell data were normalized to FBG0/LN100 sample. Error bars indicate ± SEM. Asterisk denotes p
    Dmem F12, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 71349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dmem f12
    Growth rate and morphology of MPM cells ( A ) HMCs; ( B ) SV40-Tag cells; ( C ) MPP89 cells; ( D ) MSTO-211H cells; ( E ) IST-MES2 cells. ( F ) Growth kinetics of HMCs and MPM cells cultured in serum-free RPMI or serum-free <t>DMEM-F12</t> (MPP89) medium for 72 h; cell number was assessed by measuring crystal violet uptake at 590 nm absorbance as described in Materials and Methods. Pictures were taken with a Leica phase contrast microscope as described in Materials and Methods.
    Dmem F12, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7585 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher advanced dmem f12
    Hepatocyte differentiation reduces the malignant potential of IHCC cells in vitro and in vivo . ( a ) Schedule for hepatocyte differentiation and counting of organoids and spheres. IHCC organoids were cultured in DM or EM for 12 days, then reseeded at 1.0 × 10 3 cells and cultured in EM or Advanced <t>DMEM/F12</t> (AdDF) with 10% FBS for 10 days. The numbers of organoids and spheres were counted. Scale bars: 500 μl. ( b ) Western blotting of the tumor initiating markers CD44, CD133 and LGR5 in IHCC organoids cultured in EM or DM (upper). Immunohistochemical staining of CD44 in IHCC organoids cultured in EM or DM (lower). Scale bars: 50 μm. ( c ) Tumor volumes of xenografted IHCC organoids cultured in EM or DM. We implanted 1 × 10 6 cells of IHCC organoids cultured in EM or DM subcutaneously into the backs of SCID mice. Tumor volumes of xenografted IHCC organoids cultured in EM (n = 8) or DM (n = 8) on SCID mice were measured.
    Advanced Dmem F12, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1284 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare dmem f12
    Morphological observation. Apoptotic cells were characterized by plasma membrane blebbing, cell shrinkage and nuclei condensingand. A. Phase-contrast photomicrographs of AF cells cultured in serum-free <t>DMEM/F12</t> with 0 to 120 μg/mL levofloxacin
    Dmem F12, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 2549 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Corning Life Sciences dmem f12
    Morphological alterations in mouse pMAC s following exposure to TCM , <t>DMEM</t> <t>/F12,</t> or LPS for 48 hrs. ( A ) DMEM /F12‐treated pMAC s showed no signs of activation: they had normal morphology with a regular round shape, abundant clear cytoplasm and ample intercellular spaces. ( B ) TCM ‐treated pMAC s had a relatively moderate activation/immune response: they exhibited obvious morphological changes with a polyhedron shape, large and sufficient pseudopodia, abundant granules in the cytoplasm, narrow intercellular spaces and densely populated features. ( C ) LPS (0.5 μg/ml)‐treated pMAC s showed excessive activation: they presented with irregular, doublet or multiple shapes and ultimately underwent cell death, which was characterized by cell membrane blebbing, cell body atrophy and nuclear condensation or fragmentation.
    Dmem F12, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 94/100, based on 914 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Mediatech dmem f12
    NeuT oncogene transformation enhanced ALA-induced PpIX fluorescence ( A ) Fluorescence spectra of MCF10A vector and NeuT cell lysates and PpIX standard (25 ng/mL in DMSO). Both vector and NeuT cells were incubated with 1 mM ALA in serum free medium for 4 h and lysed. Fluorescence spectra of cell lysates and PpIX standard were obtained using 400 ± 2.5 nm excitation. ( B – D ) Flow cytometer analysis showing forward scatter (FSC) (B), basal cell fluorescence without ALA (C), and a dose-dependent fluorescence increase after ALA incubation (D) in MCF10A vector and NeuT cells. Cells were cultured in serum free <t>DMEM/F12</t> medium with or without ALA for 4 h and cell fluorescence was measured with flow cytometry. ALA-induced fluorescence increase, calculated by subtracting basal cell fluorescence without ALA from cell fluorescence after incubation with different doses of ALA, was fit with the Michaelis-Menten enzyme kinetics. Data are presented as mean ± SD from at least 3 experiments. *** p
    Dmem F12, supplied by Mediatech, used in various techniques. Bioz Stars score: 92/100, based on 674 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The effect of solution added TGF-β in DMEM/F12/10% FBS media + DMSO on CSPG production. CSPG staining per cell data were normalized to FBG0/LN100 sample. Error bars indicate ± SEM. Asterisk denotes p

    Journal: Acta biomaterialia

    Article Title: Astrocytes Specifically Remove Surface-Adsorbed Fibrinogen and Locally Express Chondroitin Sulfate Proteoglycans

    doi: 10.1016/j.actbio.2013.02.047

    Figure Lengend Snippet: The effect of solution added TGF-β in DMEM/F12/10% FBS media + DMSO on CSPG production. CSPG staining per cell data were normalized to FBG0/LN100 sample. Error bars indicate ± SEM. Asterisk denotes p

    Article Snippet: Cells were maintained in DMEM/F12 (Gibco) supplemented with 10% fetal bovine serum (FBS, Sigma).

    Techniques: Staining

    Sphere formation and differentiation from the sciatic nerve in suspension culture. (A) The sciatic nerve was cultured in DMEM-F12 medium containing bFGF. The ND-GFP-expressing cells proliferated and formed spheres by day 32. Bar: 100 µm. (B) The spheres expressing ND-GFP co-expressed p75 NTR and CD34 but did not express β-III tubulin S100 and GFAP. Bar: 100 µm. (C) The ND-GFP-expressing spheres were switched to RPMI 1640 medium containing 10% FBS from DMEM-F12 containing B-27, N2 and bFGF and began to differentiate. At 7 days after switching into medium containing FBS, β-III tubulin-positive neuronal cells which expressed ND-GFP were observed. Bar: 10 µm. (D) At 14 days, the ND-GFP-expressing cells differentiated to GFAP-positive glial cells. Bar: 10 µm (E) At 7 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to K15-positive keratinocytes, some of which still expressed nestin. Bar: 10 µm. (F) At 30 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to α-SMA-positive smooth muscle cells. Bar: 50 µm.

    Journal: PLoS ONE

    Article Title: Nestin-Expressing Stem Cells Promote Nerve Growth in Long-Term 3-Dimensional Gelfoam(R)-Supported Histoculture

    doi: 10.1371/journal.pone.0067153

    Figure Lengend Snippet: Sphere formation and differentiation from the sciatic nerve in suspension culture. (A) The sciatic nerve was cultured in DMEM-F12 medium containing bFGF. The ND-GFP-expressing cells proliferated and formed spheres by day 32. Bar: 100 µm. (B) The spheres expressing ND-GFP co-expressed p75 NTR and CD34 but did not express β-III tubulin S100 and GFAP. Bar: 100 µm. (C) The ND-GFP-expressing spheres were switched to RPMI 1640 medium containing 10% FBS from DMEM-F12 containing B-27, N2 and bFGF and began to differentiate. At 7 days after switching into medium containing FBS, β-III tubulin-positive neuronal cells which expressed ND-GFP were observed. Bar: 10 µm. (D) At 14 days, the ND-GFP-expressing cells differentiated to GFAP-positive glial cells. Bar: 10 µm (E) At 7 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to K15-positive keratinocytes, some of which still expressed nestin. Bar: 10 µm. (F) At 30 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to α-SMA-positive smooth muscle cells. Bar: 50 µm.

    Article Snippet: Culture medium used was DMEM-F12 medium (GIBCO/BRL) containing B-27 (2.5%) (GIBCO/BRL), N2 (1%) (GIBCO/BRL) and 1% penicillin and streptomycin (GIBCO/BRL).

    Techniques: Cell Culture, Expressing

    IL-12 p40 induces the activation of NF- κ B in BV-2 glial cells A , cells incubated in serum-free DMEM/F-12 were treated with different concentrations of p40 2 .” B, lanes 1–3 represent nuclear extract of control cells, nuclear extract of p40 2 -treated cells, and nuclear extract of p40 2 -treated cells incubated with a 100-fold excess of unlabeled oligonucleotide. The concentration of p40 2 used in this experiment was 5 ng/ml. The upper arrow indicates the induced NF- κ B band, and the lower arrow indicates the unbound probe. C , cells plated at 50–60% confluence in 6-well plates were cotransfected with 1 μ g of pNF- κ B-Luc (an NF- κ B-dependent reporter construct) and 50 ng of pRL-TK (a plasmid encoding Renilla .” After 24 h of transfection, cells were stimulated with different concentrations of p40 2 for 6 h under serum-free conditions. Firefly and Renilla .”

    Journal: The Journal of biological chemistry

    Article Title: Induction of Nitric-oxide Synthase and Activation of NF-κB by Interleukin-12 p40 in Microglial Cells *

    doi: 10.1074/jbc.M008262200

    Figure Lengend Snippet: IL-12 p40 induces the activation of NF- κ B in BV-2 glial cells A , cells incubated in serum-free DMEM/F-12 were treated with different concentrations of p40 2 .” B, lanes 1–3 represent nuclear extract of control cells, nuclear extract of p40 2 -treated cells, and nuclear extract of p40 2 -treated cells incubated with a 100-fold excess of unlabeled oligonucleotide. The concentration of p40 2 used in this experiment was 5 ng/ml. The upper arrow indicates the induced NF- κ B band, and the lower arrow indicates the unbound probe. C , cells plated at 50–60% confluence in 6-well plates were cotransfected with 1 μ g of pNF- κ B-Luc (an NF- κ B-dependent reporter construct) and 50 ng of pRL-TK (a plasmid encoding Renilla .” After 24 h of transfection, cells were stimulated with different concentrations of p40 2 for 6 h under serum-free conditions. Firefly and Renilla .”

    Article Snippet: Fetal bovine serum, Hank's balanced salt solution and DMEM/F-12 were from Life Technologies, Inc. L- N G -Monomethylarginine (L-NMMA) and D- N G -monomethylarginine (D-NMMA), NF- κ B SN50, and NF- κ B SN50 M were purchased from Biomol.

    Techniques: Activation Assay, Incubation, Concentration Assay, Construct, Plasmid Preparation, Transfection

    (A) Apoptosis of the chondrocytes was analyzed using a TUNEL assay. TNF-α was shown to induce apoptosis and DNA fragmentation in fluorescence micrographs of the rat chondrocytes in vitro (magnification, x200). Chondrocytes were treated with TNF-α (30 ng/ml) or Dulbecco's modified Eagle's medium for 24 h. Upper panel, representative images of TUNEL staining. Lower panel, representative images showing nuclear staining of DNA (DAPI). LRP1 knockdown increased the apoptosis of chondrocytes. (B) Quantitative analysis of the percentages of TNF-α-induced TUNEL-positive cells. Data are presented as the mean ± standard deviation. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Lentivirus-induced knockdown of LRP1 induces osteoarthritic-like effects and increases susceptibility to apoptosis in chondrocytes via the nuclear factor-κB pathway

    doi: 10.3892/etm.2015.2471

    Figure Lengend Snippet: (A) Apoptosis of the chondrocytes was analyzed using a TUNEL assay. TNF-α was shown to induce apoptosis and DNA fragmentation in fluorescence micrographs of the rat chondrocytes in vitro (magnification, x200). Chondrocytes were treated with TNF-α (30 ng/ml) or Dulbecco's modified Eagle's medium for 24 h. Upper panel, representative images of TUNEL staining. Lower panel, representative images showing nuclear staining of DNA (DAPI). LRP1 knockdown increased the apoptosis of chondrocytes. (B) Quantitative analysis of the percentages of TNF-α-induced TUNEL-positive cells. Data are presented as the mean ± standard deviation. *P

    Article Snippet: Dulbecco's modified Eagle's medium (DMEM)/F-12 nutrient mixture and fetal bovine serum (FBS) were purchased from Gibco Life Technologies (Carlsbad, CA, USA).

    Techniques: TUNEL Assay, Fluorescence, In Vitro, Modification, Staining, Standard Deviation

    Effects of LRP1 on the TNF-α-induced IκB kinase-nuclear factor-κB signaling pathway in chondrocytes. Equal quantities of cell extract were subjected to western blot analysis for IκB and NF-κB subunit p65/RelA. Control and shLRP1 chondrocytes were starved in Dulbecco's modified Eagle's medium overnight, and treated with a NF-κB inhibitor (Bay 11–7082; 10 µM) for 30 min (A) without TNF-α stimulation and (B) followed by stimulation with TNF-α (30 ng/ml) for 30 min. (C and D) mRNA and (E) protein expression levels of iNOS were assessed using quantitative polymerase chain reaction and western blot analysis, respectively. **P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Lentivirus-induced knockdown of LRP1 induces osteoarthritic-like effects and increases susceptibility to apoptosis in chondrocytes via the nuclear factor-κB pathway

    doi: 10.3892/etm.2015.2471

    Figure Lengend Snippet: Effects of LRP1 on the TNF-α-induced IκB kinase-nuclear factor-κB signaling pathway in chondrocytes. Equal quantities of cell extract were subjected to western blot analysis for IκB and NF-κB subunit p65/RelA. Control and shLRP1 chondrocytes were starved in Dulbecco's modified Eagle's medium overnight, and treated with a NF-κB inhibitor (Bay 11–7082; 10 µM) for 30 min (A) without TNF-α stimulation and (B) followed by stimulation with TNF-α (30 ng/ml) for 30 min. (C and D) mRNA and (E) protein expression levels of iNOS were assessed using quantitative polymerase chain reaction and western blot analysis, respectively. **P

    Article Snippet: Dulbecco's modified Eagle's medium (DMEM)/F-12 nutrient mixture and fetal bovine serum (FBS) were purchased from Gibco Life Technologies (Carlsbad, CA, USA).

    Techniques: Western Blot, Modification, Expressing, Real-time Polymerase Chain Reaction

    LRP1 knockdown inhibits TNF-α-induced activation of the phosphoinositide 3-kinase/Akt signaling pathway in chondrocytes. Following starvation in Dulbecco's modified Eagle's medium overnight, chondrocytes were treated with or without TNF-α (30 ng/ml) for 2 h. Densitometric analyses of the bands were conducted by computerized laser densitometry and normalized against GAPDH. (A) Protein levels of total Akt, p-Akt and GAPDH were analyzed using western blot analysis. Values are expressed as the p-Akt/Akt ratio. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Lentivirus-induced knockdown of LRP1 induces osteoarthritic-like effects and increases susceptibility to apoptosis in chondrocytes via the nuclear factor-κB pathway

    doi: 10.3892/etm.2015.2471

    Figure Lengend Snippet: LRP1 knockdown inhibits TNF-α-induced activation of the phosphoinositide 3-kinase/Akt signaling pathway in chondrocytes. Following starvation in Dulbecco's modified Eagle's medium overnight, chondrocytes were treated with or without TNF-α (30 ng/ml) for 2 h. Densitometric analyses of the bands were conducted by computerized laser densitometry and normalized against GAPDH. (A) Protein levels of total Akt, p-Akt and GAPDH were analyzed using western blot analysis. Values are expressed as the p-Akt/Akt ratio. *P

    Article Snippet: Dulbecco's modified Eagle's medium (DMEM)/F-12 nutrient mixture and fetal bovine serum (FBS) were purchased from Gibco Life Technologies (Carlsbad, CA, USA).

    Techniques: Activation Assay, Modification, Western Blot

    Rod-shaped crystals develop in CHO cells overexpressing recombinant human IgG. A and B , suspension-cultured CHO cells were withdrawn from a 50-liter bioreactor bag on day 5 and immobilized on 3-triethoxysilylpropylamine-coated glass coverslips. Cells were paraformaldehyde-fixed and visualized using a DIC microscope. Scale bars , 50 μm. C , suspension-cultured CHO cells were seeded onto polylysine-coated coverslips and statically cultured for 24 h using DMEM/F-12 medium supplemented with 10% FBS in a humidified incubator at 37 °C, 5% CO 2 . Cells were paraformaldehyde-fixed and visualized using a DIC microscope. Scale bar , 20 or 50 μm as indicated.

    Journal: The Journal of Biological Chemistry

    Article Title: In Vivo

    doi: 10.1074/jbc.M110.204362

    Figure Lengend Snippet: Rod-shaped crystals develop in CHO cells overexpressing recombinant human IgG. A and B , suspension-cultured CHO cells were withdrawn from a 50-liter bioreactor bag on day 5 and immobilized on 3-triethoxysilylpropylamine-coated glass coverslips. Cells were paraformaldehyde-fixed and visualized using a DIC microscope. Scale bars , 50 μm. C , suspension-cultured CHO cells were seeded onto polylysine-coated coverslips and statically cultured for 24 h using DMEM/F-12 medium supplemented with 10% FBS in a humidified incubator at 37 °C, 5% CO 2 . Cells were paraformaldehyde-fixed and visualized using a DIC microscope. Scale bar , 20 or 50 μm as indicated.

    Article Snippet: Suspension-cultured CHO cells were withdrawn from the CultiBag via a sampling portal on day 5 or 6 and seeded onto polylysine-coated glass coverslips using DMEM/F-12 medium (Invitrogen) supplemented with 10% FBS.

    Techniques: Recombinant, Cell Culture, Microscopy

    Overexpression of AIB1 localizes AIB1-Δ4 to the nucleus. a , CHO cells were transfected with FLAG AIB1-Δ4 alone or with an equal amount of HA-AIB1 and plated on glass coverslips in DMEM F-12 + 10% FBS. Cells were fixed and permeabilized

    Journal: The Journal of Biological Chemistry

    Article Title: Role of the Nuclear Receptor Coactivator AIB1-?4 Splice Variant in the Control of Gene Transcription *

    doi: 10.1074/jbc.M110.216200

    Figure Lengend Snippet: Overexpression of AIB1 localizes AIB1-Δ4 to the nucleus. a , CHO cells were transfected with FLAG AIB1-Δ4 alone or with an equal amount of HA-AIB1 and plated on glass coverslips in DMEM F-12 + 10% FBS. Cells were fixed and permeabilized

    Article Snippet: CHO cells were grown in DMEM F-12 (Invitrogen) with 10% FBS.

    Techniques: Over Expression, Transfection

    Effects of Mito-ChM on basal OCR and bioenergetics functions in MCF-7 and MCF-10A cells. ( A ) Experimental protocol for bioenergetic functional assay. To determine the mitochondrial and glycolytic function of MCF-7 and MCF-10A cells in response to Mito-ChM (1–10 μM), we used the bioenergetic functional assay previously described (4,25). After seeding and treatment, MCF-7 cells and MCF-10A cells were subsequently washed with complete media (MEM-α for MCF-7 and DMEM/F12 for MCF-10A) and either assayed immediately, or returned to a 37°C incubator for 24, 48, or 72 h. The relative time of treatment and post-treatment incubation that corresponds to the appropriate figures is indicated. ( B ) MCF-7 and MCF-10A cells were assayed for OCR immediately after treatment with Mito-ChM (1–10 μM) for 4 h, ( C ) after incubation without Mito-ChM for an additional 24 h, ( D ) after additional incubation without Mito-ChM for 48 h, and ( E ) after additional incubation without Mito-ChM for 72 h.

    Journal: BMC Cancer

    Article Title: Mitochondria-targeted vitamin E analogs inhibit breast cancer cell energy metabolism and promote cell death

    doi: 10.1186/1471-2407-13-285

    Figure Lengend Snippet: Effects of Mito-ChM on basal OCR and bioenergetics functions in MCF-7 and MCF-10A cells. ( A ) Experimental protocol for bioenergetic functional assay. To determine the mitochondrial and glycolytic function of MCF-7 and MCF-10A cells in response to Mito-ChM (1–10 μM), we used the bioenergetic functional assay previously described (4,25). After seeding and treatment, MCF-7 cells and MCF-10A cells were subsequently washed with complete media (MEM-α for MCF-7 and DMEM/F12 for MCF-10A) and either assayed immediately, or returned to a 37°C incubator for 24, 48, or 72 h. The relative time of treatment and post-treatment incubation that corresponds to the appropriate figures is indicated. ( B ) MCF-7 and MCF-10A cells were assayed for OCR immediately after treatment with Mito-ChM (1–10 μM) for 4 h, ( C ) after incubation without Mito-ChM for an additional 24 h, ( D ) after additional incubation without Mito-ChM for 48 h, and ( E ) after additional incubation without Mito-ChM for 72 h.

    Article Snippet: MCF-10A cells were cultured in DMEM/F12 media (1:1) (Invitrogen) supplemented with 5% horse serum, bovine insulin (10 μg/ml), epidermal growth factor (20 ng/ml), cholera toxin (100 ng/ml), and hydrocortisone (0.5 μg/ml), penicillin (100 U/ml) and streptomycin (100 μg/ml).

    Techniques: Functional Assay, Incubation

    Schematic representation of hTSC differentiation into hOLCs. hTSCs; Human theca stem cells, hOLCs; Human oocyte like cells, DMEM/F12; Dulbecco’s Modified Eagle’s medium F12, FBS; Fetal bovine serum, BSA; Bovine serum albumin, FSH; Follicle-stimulating hormone, and LH; Luteinizing hormone.

    Journal: Cell Journal (Yakhteh)

    Article Title: Human Ovarian Theca-Derived Multipotent Stem Cells Have The Potential to Differentiate into Oocyte-Like Cells In Vitro

    doi: 10.22074/cellj.2019.5651

    Figure Lengend Snippet: Schematic representation of hTSC differentiation into hOLCs. hTSCs; Human theca stem cells, hOLCs; Human oocyte like cells, DMEM/F12; Dulbecco’s Modified Eagle’s medium F12, FBS; Fetal bovine serum, BSA; Bovine serum albumin, FSH; Follicle-stimulating hormone, and LH; Luteinizing hormone.

    Article Snippet: Follicles were punctured in Dulbecco’s Modified Eagle’s medium F12 (DMEM/F12, Gibco, Grand Island, NY, USA).

    Techniques: Modification

    EML4-ALK fusion promotes hormone-independent growth and confers resistance to fulvestrant. (A) EML4-ALK fusion promoted hormone independent growth of T47D cells. T47D cells with empty vector (EV) or EML4-ALK fusion were seeded into 96-well plates in Dulbecco’s Modified Eagle Medium (DMEM)/F12 medium supplemented with either 10% fetal bovine serum (FBS) or 10% charcoal stripped bovine serum (CSS), and cell proliferation was assayed with resazurin. (B) Western blot analysis of phosphorylated AKT, ERK and PLCγ in T47D cells expressing EML4-ALK fusion that were cultured with either 10% FBS or 10% CSS and treated with 100 nM ceritinib for 4 h or 24 h. (C) Proliferative inhibition of T47D cells expressing EML4-ALK fusion by ceritinib. T47D cells with empty vector (EV) or EML4-ALK fusion were treated with various doses of ceritinib in medium supplemented with 10% FBS or 10% CSS. (D) EML4-ALK fusion confers resistance to fulvestrant. T47D cells with EV or EML4-ALK fusion seeded into 96-well plates were treated with 100 nM fulvestrant (ICI) and/or 100 nM ceritinib.

    Journal: Annals of oncology : official journal of the European Society for Medical Oncology

    Article Title: Enrichment of kinase fusions in ESR1 wild-type, metastatic breast cancer revealed by a systematic analysis of 4854 patients

    doi: 10.1016/j.annonc.2020.04.008

    Figure Lengend Snippet: EML4-ALK fusion promotes hormone-independent growth and confers resistance to fulvestrant. (A) EML4-ALK fusion promoted hormone independent growth of T47D cells. T47D cells with empty vector (EV) or EML4-ALK fusion were seeded into 96-well plates in Dulbecco’s Modified Eagle Medium (DMEM)/F12 medium supplemented with either 10% fetal bovine serum (FBS) or 10% charcoal stripped bovine serum (CSS), and cell proliferation was assayed with resazurin. (B) Western blot analysis of phosphorylated AKT, ERK and PLCγ in T47D cells expressing EML4-ALK fusion that were cultured with either 10% FBS or 10% CSS and treated with 100 nM ceritinib for 4 h or 24 h. (C) Proliferative inhibition of T47D cells expressing EML4-ALK fusion by ceritinib. T47D cells with empty vector (EV) or EML4-ALK fusion were treated with various doses of ceritinib in medium supplemented with 10% FBS or 10% CSS. (D) EML4-ALK fusion confers resistance to fulvestrant. T47D cells with EV or EML4-ALK fusion seeded into 96-well plates were treated with 100 nM fulvestrant (ICI) and/or 100 nM ceritinib.

    Article Snippet: MCF7 and T47D cells were grown in Dulbecco’s Modified Eagle Medium (DMEM)/F12 and RPMI-1640, respectively, supplemented with 10% fetal bovine serum (FBS) or 10% charcoal stripped bovine serum (Gibco, Waltham, MA), 100 μg/ml penicillin, 100 mg/ml streptomycin (Corning, Corning, NY), and 4 mM glutamine.

    Techniques: Plasmid Preparation, Modification, Western Blot, Expressing, Cell Culture, Inhibition

    LMNA-NTRK1 fusion promotes hormone-independent growth and confers resistance to endocrine therapy. (A) Western blot analysis of ERα, PgR and phosphorylation of AKT, ERK and PLCγ in MCF7 cells expressing LMNA-NTRK1 fusion that were cultured with either 10% FBS or 10% CSS and treated with 100 nM larotrectinib for 4 h or 24 h. (B) LMNA-NTRK1 fusion promoted hormone-independent growth of MCF7 cells. MCF7 cells with empty vector (EV) or LMNA-NTRK1 fusion were seeded into 96-well plates in DMEM/F12 medium supplemented with either 10% FBS or 10% CSS, and cell proliferation was assayed with resazurin. (C) Proliferative inhibition of MCF7 cells expressing LMNA-NTRK1 fusion by larotrectinib. MCF7 cells with EV or LMNA-NTRK1 fusion were treated with various doses of larotrectinib in medium supplemented with 10% FBS or 10% CSS. (D) LMNA-NTRK1 fusion confers resistance to fulvestrant. MCF7 cells with EV or LMNA-NTRK1 fusion seeded into 96-well plates were treated with various doses of fulvestrant (ICI) and/or 100 nM larotrectinib. (E) LMNA-NTRKl-expressing MCF7 xenografts after estrogen deprivation. MCF7 xenograft tumors expressing LMNA-NTRK1 fusion were established in nude mice. Then the estradiol pellets were removed, and the mice were treated with either vehicle or 200 mg/kg of larotrectinib via gavage once daily for 2 weeks. The mice bearing MCF7 tumors expressing the EV were treated with vehicle as control. The relative tumor volume compared with the volume measured before treatment of each group ± standard deviation ( n = 10 mice/group) is shown with t-test P values. CSS, charcoal-stripped bovine serum; DMEM, Dulbecco’s Modified Eagle Medium; DMSO, dimethyl sulfoxide; ERα, estrogen receptor alpha; FBS, fetal bovine serum; PgR, progesterone receptor.

    Journal: Annals of oncology : official journal of the European Society for Medical Oncology

    Article Title: Enrichment of kinase fusions in ESR1 wild-type, metastatic breast cancer revealed by a systematic analysis of 4854 patients

    doi: 10.1016/j.annonc.2020.04.008

    Figure Lengend Snippet: LMNA-NTRK1 fusion promotes hormone-independent growth and confers resistance to endocrine therapy. (A) Western blot analysis of ERα, PgR and phosphorylation of AKT, ERK and PLCγ in MCF7 cells expressing LMNA-NTRK1 fusion that were cultured with either 10% FBS or 10% CSS and treated with 100 nM larotrectinib for 4 h or 24 h. (B) LMNA-NTRK1 fusion promoted hormone-independent growth of MCF7 cells. MCF7 cells with empty vector (EV) or LMNA-NTRK1 fusion were seeded into 96-well plates in DMEM/F12 medium supplemented with either 10% FBS or 10% CSS, and cell proliferation was assayed with resazurin. (C) Proliferative inhibition of MCF7 cells expressing LMNA-NTRK1 fusion by larotrectinib. MCF7 cells with EV or LMNA-NTRK1 fusion were treated with various doses of larotrectinib in medium supplemented with 10% FBS or 10% CSS. (D) LMNA-NTRK1 fusion confers resistance to fulvestrant. MCF7 cells with EV or LMNA-NTRK1 fusion seeded into 96-well plates were treated with various doses of fulvestrant (ICI) and/or 100 nM larotrectinib. (E) LMNA-NTRKl-expressing MCF7 xenografts after estrogen deprivation. MCF7 xenograft tumors expressing LMNA-NTRK1 fusion were established in nude mice. Then the estradiol pellets were removed, and the mice were treated with either vehicle or 200 mg/kg of larotrectinib via gavage once daily for 2 weeks. The mice bearing MCF7 tumors expressing the EV were treated with vehicle as control. The relative tumor volume compared with the volume measured before treatment of each group ± standard deviation ( n = 10 mice/group) is shown with t-test P values. CSS, charcoal-stripped bovine serum; DMEM, Dulbecco’s Modified Eagle Medium; DMSO, dimethyl sulfoxide; ERα, estrogen receptor alpha; FBS, fetal bovine serum; PgR, progesterone receptor.

    Article Snippet: MCF7 and T47D cells were grown in Dulbecco’s Modified Eagle Medium (DMEM)/F12 and RPMI-1640, respectively, supplemented with 10% fetal bovine serum (FBS) or 10% charcoal stripped bovine serum (Gibco, Waltham, MA), 100 μg/ml penicillin, 100 mg/ml streptomycin (Corning, Corning, NY), and 4 mM glutamine.

    Techniques: Western Blot, Expressing, Cell Culture, Plasmid Preparation, Inhibition, Mouse Assay, Standard Deviation, Modification

    Endothelial cells increase the malignant potential of liver cancer cells. (A) A schematic illustration showing the procedure for the acquisition of the conditioned medium obtained from HUVECs and the applications in this study. (B) Morphological changes of HepG2 cells after the treatment with ECM. (C) Effect of ECM on the growth of HepG2 cells. After HepG2 cells were treated with ECM, the cell counting was performed at 24, 48 and 72 h. Increased (D) migration and (E) invasion of HepG2 cells after the treatment with ECM. The migratory and invasive properties of the cells were measured using the Transwell chamber. (F) Western blot analysis for the expressions of N-cadherin and vimentin after the treatment with ECM. Experiments were performed in triplicate, and the data shown are representative of a typical experiment. (G) Quantification of sphere-forming abilities of HepG2 cells after the treatment with ECM. The cells were grown in DMEM/F12 supplemented with B27, N2, basic fibroblast- and epidermal growth factor onto 24-well ultra-low attachment plates at 300 cells per well for 7 days, and the size of spheres were determined. To measure the size of sphere, 12 spheres per group (n=12/group) were randomly selected. The average size of each sphere is quantified in the standard deviation and shown in the representative graph. Results from three independent experiments are expressed as mean ± 1 SEM (*P

    Journal: Oncology Letters

    Article Title: Irradiated endothelial cells modulate the malignancy of liver cancer cells

    doi: 10.3892/ol.2018.9833

    Figure Lengend Snippet: Endothelial cells increase the malignant potential of liver cancer cells. (A) A schematic illustration showing the procedure for the acquisition of the conditioned medium obtained from HUVECs and the applications in this study. (B) Morphological changes of HepG2 cells after the treatment with ECM. (C) Effect of ECM on the growth of HepG2 cells. After HepG2 cells were treated with ECM, the cell counting was performed at 24, 48 and 72 h. Increased (D) migration and (E) invasion of HepG2 cells after the treatment with ECM. The migratory and invasive properties of the cells were measured using the Transwell chamber. (F) Western blot analysis for the expressions of N-cadherin and vimentin after the treatment with ECM. Experiments were performed in triplicate, and the data shown are representative of a typical experiment. (G) Quantification of sphere-forming abilities of HepG2 cells after the treatment with ECM. The cells were grown in DMEM/F12 supplemented with B27, N2, basic fibroblast- and epidermal growth factor onto 24-well ultra-low attachment plates at 300 cells per well for 7 days, and the size of spheres were determined. To measure the size of sphere, 12 spheres per group (n=12/group) were randomly selected. The average size of each sphere is quantified in the standard deviation and shown in the representative graph. Results from three independent experiments are expressed as mean ± 1 SEM (*P

    Article Snippet: Sphere forming assay Cells were grown in serum-free DMEM/F12 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with B27 (Gibco; Thermo Fisher Scientific, Inc.), N2 (Gibco; Thermo Fisher Scientific, Inc.), 20 ng/ml basic fibroblast (Peprotech, London, UK) and 20 ng/ml epidermal growth factor (Peprotech) onto 24-well ultra-low attachment plates at 300 cells per well for 7 or 14 days, and then the size and number of spheres were determined using a phase-contrast Nikon microscope (TS100; Nikon).

    Techniques: Cell Counting, Migration, Western Blot, Standard Deviation

    2 Gy-irradiated endothelial cells enhance the malignant potential of liver cancer cells. (A) Migratory and (B) invasive properties of HepG2 cells after the treatment with ECM obtained from irradiated HUVECs. These properties of the cells were measured using the Transwell chamber (×200 magnification). (C) Western blot analysis for the expressions of E-cadherin, N-cadherin, vimentin, slug, zeb1 and snail after the treatment with ECM. Experiments were performed in triplicate, and the data shown are representative of a typical experiment. (D) Quantification of sphere-forming abilities of HepG2 cells after the treatment with ECM obtained from irradiated HUVECs. The cells were grown in DMEM/F12 supplemented with B27, N2, basic fibroblast- and epidermal growth factor onto 24-well ultra-low attachment plates at 300 cells per well for 7 days, and the size of spheres were determined. To measure the size of sphere, 12 spheres per group (n=12/group) were randomly selected. The average size of each sphere is quantified in the standard deviation and shown in the representative graph. *P

    Journal: Oncology Letters

    Article Title: Irradiated endothelial cells modulate the malignancy of liver cancer cells

    doi: 10.3892/ol.2018.9833

    Figure Lengend Snippet: 2 Gy-irradiated endothelial cells enhance the malignant potential of liver cancer cells. (A) Migratory and (B) invasive properties of HepG2 cells after the treatment with ECM obtained from irradiated HUVECs. These properties of the cells were measured using the Transwell chamber (×200 magnification). (C) Western blot analysis for the expressions of E-cadherin, N-cadherin, vimentin, slug, zeb1 and snail after the treatment with ECM. Experiments were performed in triplicate, and the data shown are representative of a typical experiment. (D) Quantification of sphere-forming abilities of HepG2 cells after the treatment with ECM obtained from irradiated HUVECs. The cells were grown in DMEM/F12 supplemented with B27, N2, basic fibroblast- and epidermal growth factor onto 24-well ultra-low attachment plates at 300 cells per well for 7 days, and the size of spheres were determined. To measure the size of sphere, 12 spheres per group (n=12/group) were randomly selected. The average size of each sphere is quantified in the standard deviation and shown in the representative graph. *P

    Article Snippet: Sphere forming assay Cells were grown in serum-free DMEM/F12 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with B27 (Gibco; Thermo Fisher Scientific, Inc.), N2 (Gibco; Thermo Fisher Scientific, Inc.), 20 ng/ml basic fibroblast (Peprotech, London, UK) and 20 ng/ml epidermal growth factor (Peprotech) onto 24-well ultra-low attachment plates at 300 cells per well for 7 or 14 days, and then the size and number of spheres were determined using a phase-contrast Nikon microscope (TS100; Nikon).

    Techniques: Irradiation, Western Blot, Standard Deviation

    Morphology of parent OVA-BS4 (magnification 200X) ( a ) and OVA-BS4 spheroids (magnification 100X) ( b ). Parent OVA-BS4 were cultured in RPMI 10%FBS in adhesion condition, whereas OVA-BS4 spheroids were obtained from parent OVA-BS4 cell line under selective culture condition (serum-free DMEM/F12 medium supplemented with 5 μg/ml human insulin, 20 ng/ml EGF, 10 ng/ml bFGF, and B27 Supplement, in ultra low-attachment plates)

    Journal: BMC Cancer

    Article Title: MAL gene overexpression as a marker of high-grade serous ovarian carcinoma stem-like cells that predicts chemoresistance and poor prognosis

    doi: 10.1186/s12885-017-3334-1

    Figure Lengend Snippet: Morphology of parent OVA-BS4 (magnification 200X) ( a ) and OVA-BS4 spheroids (magnification 100X) ( b ). Parent OVA-BS4 were cultured in RPMI 10%FBS in adhesion condition, whereas OVA-BS4 spheroids were obtained from parent OVA-BS4 cell line under selective culture condition (serum-free DMEM/F12 medium supplemented with 5 μg/ml human insulin, 20 ng/ml EGF, 10 ng/ml bFGF, and B27 Supplement, in ultra low-attachment plates)

    Article Snippet: In detail, parent OVA-BS4 cells were trypsinized and placed at a density of 5 × 105 /ml in ultra-low attachment plates (Corning, New York, USA), in serum-free DMEM/F12 medium (Gibco, Life Technologies, Carlsbad, California, USA) supplemented with 5 μg/ml human insulin (Sigma, St. Louis, Missouri, USA), 20 ng/ml human recombinant epidermal growth factor (EGF, Gibco), 10 ng/ml human recombinant basic fibroblast growth factor (bFGF, Gibco) and B27 Supplement (Gibco) [ ].

    Techniques: Cell Culture

    General scheme of ESC differentiation and experimental outline of study. a After two passages on gelatin, ESC differentiation started with EB formation, followed by selection of Nestin + cells in ITSFn medium and subsequent expansion in DMEM/F12 + B27 medium in the presence of bFGF. On day 24 of differentiation, cells were purified by FACS and MACS to yield a population of SSEA-1 – /PSA-NCAM + progenitors, which were labeled with BrdU for 48 hours prior to transplantation. b Transplantation was performed on DPO7, the surgery for biotin dextran tracing took place on DPO21, and the von Frey filament test immediately before perfusion on DPO42. Motor behavior by BMS scoring was tested 1 day prior to SCI (DPO–1) and on DPO1, 6, 8, 10, 14, 21, 28, 35, and 42. bFGF basic fibroblast growth factor, BrdU 5-bromo-2′-deoxyuridine, DMEM Dulbecco’s modified Eagle’s medium, DPO days post operation, EB embryoid body, ESC embryonic stem cell, FACS , fluorescence-activated cell sorting, ITSFn insulin, transferrin, selenium chloride, fibronectin, MACS magnetic-activated cell sorting, PSA-NCAM polysialylated neural cell adhesion molecule, SCI spinal cord injury, SSEA-1 stage-specific embryonic antigen-1

    Journal: Stem Cell Research & Therapy

    Article Title: PSA-NCAM positive neural progenitors stably expressing BDNF promote functional recovery in a mouse model of spinal cord injury

    doi: 10.1186/s13287-015-0268-x

    Figure Lengend Snippet: General scheme of ESC differentiation and experimental outline of study. a After two passages on gelatin, ESC differentiation started with EB formation, followed by selection of Nestin + cells in ITSFn medium and subsequent expansion in DMEM/F12 + B27 medium in the presence of bFGF. On day 24 of differentiation, cells were purified by FACS and MACS to yield a population of SSEA-1 – /PSA-NCAM + progenitors, which were labeled with BrdU for 48 hours prior to transplantation. b Transplantation was performed on DPO7, the surgery for biotin dextran tracing took place on DPO21, and the von Frey filament test immediately before perfusion on DPO42. Motor behavior by BMS scoring was tested 1 day prior to SCI (DPO–1) and on DPO1, 6, 8, 10, 14, 21, 28, 35, and 42. bFGF basic fibroblast growth factor, BrdU 5-bromo-2′-deoxyuridine, DMEM Dulbecco’s modified Eagle’s medium, DPO days post operation, EB embryoid body, ESC embryonic stem cell, FACS , fluorescence-activated cell sorting, ITSFn insulin, transferrin, selenium chloride, fibronectin, MACS magnetic-activated cell sorting, PSA-NCAM polysialylated neural cell adhesion molecule, SCI spinal cord injury, SSEA-1 stage-specific embryonic antigen-1

    Article Snippet: After 8 days, cells were trypsinized and replated onto dishes coated with poly-ornithin/laminin (Sigma Aldrich, St. Louis, MO, USA) at a density of 1.5 × 105 cells/cm2 in DMEM/F12/Glutamax supplemented with B27 (all from Invitrogen, Carlsbad, CA, USA) and 10 ng/ml basic fibroblast growth factor (bFGF; Sigma Aldrich).

    Techniques: Selection, Purification, FACS, Magnetic Cell Separation, Labeling, Transplantation Assay, Modification, Fluorescence

    Morphology recovery of HCE cells at passage 101. A : Control HCE cells cultured in 20% FBS-containing DMEM/F12 (1:1) medium, showing the fibroblastic morphology. B : HCE cells cultured in 5% BCS-containing MEM medium, showing the more corneal endotheloid-like morphology. Scale bar: 50 μm.

    Journal: Molecular Vision

    Article Title: Establishment of a continuous untransfected human corneal endothelial cell line and its biocompatibility to denuded amniotic membrane

    doi:

    Figure Lengend Snippet: Morphology recovery of HCE cells at passage 101. A : Control HCE cells cultured in 20% FBS-containing DMEM/F12 (1:1) medium, showing the fibroblastic morphology. B : HCE cells cultured in 5% BCS-containing MEM medium, showing the more corneal endotheloid-like morphology. Scale bar: 50 μm.

    Article Snippet: Morphology recovery Morphology recovery of HCE cells at passage 101 was performed with 5%, 10%, 15% BCS-supplemented MEM, DMEM and DMEM/F12 medium (Invitrogen), respectively, as previously described [ ].

    Techniques: Cell Culture

    DCN evokes different responses in cultured human GCs: transient elevations of intracellular Ca 2+ and generation of reactive oxygen species (ROS). ( A ) A representative experiment, in which GCs were treated with medium (DMEM/Ham’s F-12; −)

    Journal: Human Reproduction (Oxford, England)

    Article Title: Decorin is a part of the ovarian extracellular matrix in primates and may act as a signaling molecule

    doi: 10.1093/humrep/des297

    Figure Lengend Snippet: DCN evokes different responses in cultured human GCs: transient elevations of intracellular Ca 2+ and generation of reactive oxygen species (ROS). ( A ) A representative experiment, in which GCs were treated with medium (DMEM/Ham’s F-12; −)

    Article Snippet: In brief, GCs from three different pools of patients (Days 2 and 3) were grown on glass cover slips for 24 h in DMEM/Ham’s F-12 supplemented with 10% FCS and 1% penicillin/streptomycin, loaded with 5 µM fluo-4, acetoxy-methylester (fluo-4 AM, Molecular Probes, Eugene, OR, USA) for 30 min at 37°C and 5% CO2 .

    Techniques: Cell Culture

    Improvement of ureteric branching in Gen1 mutants by exogenous RA in vitro. Hoxb7;Gen1 PB/PB kidney rudiments were cultured for 72 h in F12/DMEM with RA (ATRA +9-cis-RA) dissolved in DMSO (A–A′–B–B′) , and controls with DMSO only (C–C′–D–D′) . From (A) and (C) , we found that without unilateral duplex budding, supplementation with RA could obviously increase the number of branching (E) ; however, among duplex groups (B) and (D) , the branching was limited even when administrated with RA. Quantification of the total number of branching in both with and without duplex budding of metanephroi (F) . Unpaired t -test, * p = 0.05. DMEM, Dulbecco's modified Eagle's medium; DMSO, dimethyl sulfoxide; RA, retinoic acid. Color images are available online.

    Journal: DNA and Cell Biology

    Article Title: Gen1 Modulates Metanephric Morphology Through Retinoic Acid Signaling

    doi: 10.1089/dna.2018.4426

    Figure Lengend Snippet: Improvement of ureteric branching in Gen1 mutants by exogenous RA in vitro. Hoxb7;Gen1 PB/PB kidney rudiments were cultured for 72 h in F12/DMEM with RA (ATRA +9-cis-RA) dissolved in DMSO (A–A′–B–B′) , and controls with DMSO only (C–C′–D–D′) . From (A) and (C) , we found that without unilateral duplex budding, supplementation with RA could obviously increase the number of branching (E) ; however, among duplex groups (B) and (D) , the branching was limited even when administrated with RA. Quantification of the total number of branching in both with and without duplex budding of metanephroi (F) . Unpaired t -test, * p = 0.05. DMEM, Dulbecco's modified Eagle's medium; DMSO, dimethyl sulfoxide; RA, retinoic acid. Color images are available online.

    Article Snippet: E11.5 Hoxb7;Gen1PB/PB kidney rudiments were dissected from the embryos, explanted onto 0.4-mm polyester membrane Transwell supports, and cultured at the air/liquid interface for 72 h with Dulbecco's modified Eagle's medium/Nutrient Mixture F-12 (DMEM/F-12) (Gibco).

    Techniques: In Vitro, Cell Culture, Modification

    Growth rate and morphology of MPM cells ( A ) HMCs; ( B ) SV40-Tag cells; ( C ) MPP89 cells; ( D ) MSTO-211H cells; ( E ) IST-MES2 cells. ( F ) Growth kinetics of HMCs and MPM cells cultured in serum-free RPMI or serum-free DMEM-F12 (MPP89) medium for 72 h; cell number was assessed by measuring crystal violet uptake at 590 nm absorbance as described in Materials and Methods. Pictures were taken with a Leica phase contrast microscope as described in Materials and Methods.

    Journal: Oncotarget

    Article Title: P2×7 targeting inhibits growth of human mesothelioma

    doi: 10.18632/oncotarget.10430

    Figure Lengend Snippet: Growth rate and morphology of MPM cells ( A ) HMCs; ( B ) SV40-Tag cells; ( C ) MPP89 cells; ( D ) MSTO-211H cells; ( E ) IST-MES2 cells. ( F ) Growth kinetics of HMCs and MPM cells cultured in serum-free RPMI or serum-free DMEM-F12 (MPP89) medium for 72 h; cell number was assessed by measuring crystal violet uptake at 590 nm absorbance as described in Materials and Methods. Pictures were taken with a Leica phase contrast microscope as described in Materials and Methods.

    Article Snippet: MPP89 and MPP89/CytLUC cells were cultured in DMEM-F12 (Sigma-Aldrich) supplemented with 15% fetal calf serum, 100 μg/ml hygromycin Sigma-Aldrich, only MPP89/CytLUC cells), 100 U/ml penicillin and 100 mg/ml streptomycin.

    Techniques: Cell Culture, Microscopy

    Fibronectin induces phosphorylation of ER at serine-118 in MCF-7 cells through MAPK/ERK1/2. a MCF-7 cells were grown to 80% confluency and starved for 48 h in phenol red free DMEM/F12. They were then treated with vehicle (water) or FN (30 µg/ml)

    Journal: Breast cancer research and treatment

    Article Title: The tumor microenvironment modulates tamoxifen resistance in breast cancer: a role for soluble stromal factors and fibronectin through ?1 integrin

    doi: 10.1007/s10549-011-1766-x

    Figure Lengend Snippet: Fibronectin induces phosphorylation of ER at serine-118 in MCF-7 cells through MAPK/ERK1/2. a MCF-7 cells were grown to 80% confluency and starved for 48 h in phenol red free DMEM/F12. They were then treated with vehicle (water) or FN (30 µg/ml)

    Article Snippet: The MCF-7 and LM05-Mix, -E, -F cell lines were routinely maintained in growth medium, consisting of DMEM/F12 medium (Sigma-Aldrich, St. Louis, MO), supplemented with 10% fetal calf serum (FCS, GenSA, Buenos Aires, Argentina) and gentamicin, in a humidified 5% CO2 /air atmosphere.

    Techniques:

    Hepatocyte differentiation reduces the malignant potential of IHCC cells in vitro and in vivo . ( a ) Schedule for hepatocyte differentiation and counting of organoids and spheres. IHCC organoids were cultured in DM or EM for 12 days, then reseeded at 1.0 × 10 3 cells and cultured in EM or Advanced DMEM/F12 (AdDF) with 10% FBS for 10 days. The numbers of organoids and spheres were counted. Scale bars: 500 μl. ( b ) Western blotting of the tumor initiating markers CD44, CD133 and LGR5 in IHCC organoids cultured in EM or DM (upper). Immunohistochemical staining of CD44 in IHCC organoids cultured in EM or DM (lower). Scale bars: 50 μm. ( c ) Tumor volumes of xenografted IHCC organoids cultured in EM or DM. We implanted 1 × 10 6 cells of IHCC organoids cultured in EM or DM subcutaneously into the backs of SCID mice. Tumor volumes of xenografted IHCC organoids cultured in EM (n = 8) or DM (n = 8) on SCID mice were measured.

    Journal: Scientific Reports

    Article Title: Induction of differentiation of intrahepatic cholangiocarcinoma cells to functional hepatocytes using an organoid culture system

    doi: 10.1038/s41598-018-21121-6

    Figure Lengend Snippet: Hepatocyte differentiation reduces the malignant potential of IHCC cells in vitro and in vivo . ( a ) Schedule for hepatocyte differentiation and counting of organoids and spheres. IHCC organoids were cultured in DM or EM for 12 days, then reseeded at 1.0 × 10 3 cells and cultured in EM or Advanced DMEM/F12 (AdDF) with 10% FBS for 10 days. The numbers of organoids and spheres were counted. Scale bars: 500 μl. ( b ) Western blotting of the tumor initiating markers CD44, CD133 and LGR5 in IHCC organoids cultured in EM or DM (upper). Immunohistochemical staining of CD44 in IHCC organoids cultured in EM or DM (lower). Scale bars: 50 μm. ( c ) Tumor volumes of xenografted IHCC organoids cultured in EM or DM. We implanted 1 × 10 6 cells of IHCC organoids cultured in EM or DM subcutaneously into the backs of SCID mice. Tumor volumes of xenografted IHCC organoids cultured in EM (n = 8) or DM (n = 8) on SCID mice were measured.

    Article Snippet: For the organoid counting assay, organoids cultured in EM or DM were reseeded at 1.0 × 103 cells in Matrigel and then cultured in EM or Advanced DMEM/F12 with 10% FBS for 10 days.

    Techniques: In Vitro, In Vivo, Cell Culture, Western Blot, Immunohistochemistry, Staining, Mouse Assay

    Morphological observation. Apoptotic cells were characterized by plasma membrane blebbing, cell shrinkage and nuclei condensingand. A. Phase-contrast photomicrographs of AF cells cultured in serum-free DMEM/F12 with 0 to 120 μg/mL levofloxacin

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Levofloxacin increases apoptosis of rat annulus fibrosus cells via the mechanism of upregulating MMP-2 and MMP-13

    doi:

    Figure Lengend Snippet: Morphological observation. Apoptotic cells were characterized by plasma membrane blebbing, cell shrinkage and nuclei condensingand. A. Phase-contrast photomicrographs of AF cells cultured in serum-free DMEM/F12 with 0 to 120 μg/mL levofloxacin

    Article Snippet: AF cells were isolated from tissue via enzyme solution of 0.2% collagenase type II (Sigma Chemical Co., St. Louis, MD) digestion at 37°C for 1 h. The supernatant liquid was discarded after centrifugation for 5 min and digested with 0.25% trypsin/0.02% EDTA (Sigma Chemical Co., St. Louis, MD) at 37°C for 3 min, then centrifugate again and abandoned the supernatant cultured cells in complete media: DMEM/F12 (Hyclone, Logan, UT) + 10% FBS (Hyclone, Logan, UT) at 37°C in an incubator under 5% CO2 .

    Techniques: Cell Culture

    AF cell binding to collagen I. Rat AF cells were sub-cultured in six-well plates at 2×10 5 cells per well with complete culture medium. After reaching 90% confluence, the medium was changed to DMEM/F12 containing 1% FBS for 12 h to synchronize

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Levofloxacin increases apoptosis of rat annulus fibrosus cells via the mechanism of upregulating MMP-2 and MMP-13

    doi:

    Figure Lengend Snippet: AF cell binding to collagen I. Rat AF cells were sub-cultured in six-well plates at 2×10 5 cells per well with complete culture medium. After reaching 90% confluence, the medium was changed to DMEM/F12 containing 1% FBS for 12 h to synchronize

    Article Snippet: AF cells were isolated from tissue via enzyme solution of 0.2% collagenase type II (Sigma Chemical Co., St. Louis, MD) digestion at 37°C for 1 h. The supernatant liquid was discarded after centrifugation for 5 min and digested with 0.25% trypsin/0.02% EDTA (Sigma Chemical Co., St. Louis, MD) at 37°C for 3 min, then centrifugate again and abandoned the supernatant cultured cells in complete media: DMEM/F12 (Hyclone, Logan, UT) + 10% FBS (Hyclone, Logan, UT) at 37°C in an incubator under 5% CO2 .

    Techniques: Binding Assay, Cell Culture

    Changes in caspase-3 activity. Rat AF cells were sub-cultured in six-well plates at 2×10 5 cells per well with complete culture medium. After reaching 90% confluence, the medium was changed to DMEM/F12 containing 1% FBS for 12 h to synchronize

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Levofloxacin increases apoptosis of rat annulus fibrosus cells via the mechanism of upregulating MMP-2 and MMP-13

    doi:

    Figure Lengend Snippet: Changes in caspase-3 activity. Rat AF cells were sub-cultured in six-well plates at 2×10 5 cells per well with complete culture medium. After reaching 90% confluence, the medium was changed to DMEM/F12 containing 1% FBS for 12 h to synchronize

    Article Snippet: AF cells were isolated from tissue via enzyme solution of 0.2% collagenase type II (Sigma Chemical Co., St. Louis, MD) digestion at 37°C for 1 h. The supernatant liquid was discarded after centrifugation for 5 min and digested with 0.25% trypsin/0.02% EDTA (Sigma Chemical Co., St. Louis, MD) at 37°C for 3 min, then centrifugate again and abandoned the supernatant cultured cells in complete media: DMEM/F12 (Hyclone, Logan, UT) + 10% FBS (Hyclone, Logan, UT) at 37°C in an incubator under 5% CO2 .

    Techniques: Activity Assay, Cell Culture

    BD Matrigel and ECM gel, but not methylcellulose, improve GFP-SC numbers over cell:medium suspensions ( A–D ). A significant improvement in SC numbers at 12 weeks following implantation into the injured spinal cord is achieved over DMEM-F12 medium

    Journal: Journal of Neurotrauma

    Article Title: Suspension Matrices for Improved Schwann-Cell Survival after Implantation into the Injured Rat Spinal Cord

    doi: 10.1089/neu.2008.0809

    Figure Lengend Snippet: BD Matrigel and ECM gel, but not methylcellulose, improve GFP-SC numbers over cell:medium suspensions ( A–D ). A significant improvement in SC numbers at 12 weeks following implantation into the injured spinal cord is achieved over DMEM-F12 medium

    Article Snippet: For the histological studies, animals were randomly assigned to one of four treatment groups, receiving the EGFP-SC suspensions of 2 × 106 cells within DMEM/F12 medium ( n = 9, Hyclone; 8 μl total volume) or a specific fluid matrix: methylcellulose ( n = 8, Sigma-Aldrich), ECM gel ( n = 9, Sigma-Aldrich), or Matrigel ( n = 9, BD).

    Techniques:

    Morphological alterations in mouse pMAC s following exposure to TCM , DMEM /F12, or LPS for 48 hrs. ( A ) DMEM /F12‐treated pMAC s showed no signs of activation: they had normal morphology with a regular round shape, abundant clear cytoplasm and ample intercellular spaces. ( B ) TCM ‐treated pMAC s had a relatively moderate activation/immune response: they exhibited obvious morphological changes with a polyhedron shape, large and sufficient pseudopodia, abundant granules in the cytoplasm, narrow intercellular spaces and densely populated features. ( C ) LPS (0.5 μg/ml)‐treated pMAC s showed excessive activation: they presented with irregular, doublet or multiple shapes and ultimately underwent cell death, which was characterized by cell membrane blebbing, cell body atrophy and nuclear condensation or fragmentation.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: In vitro morphology, viability and cytokine secretion of uterine telocyte‐activated mouse peritoneal macrophages

    doi: 10.1111/jcmm.12711

    Figure Lengend Snippet: Morphological alterations in mouse pMAC s following exposure to TCM , DMEM /F12, or LPS for 48 hrs. ( A ) DMEM /F12‐treated pMAC s showed no signs of activation: they had normal morphology with a regular round shape, abundant clear cytoplasm and ample intercellular spaces. ( B ) TCM ‐treated pMAC s had a relatively moderate activation/immune response: they exhibited obvious morphological changes with a polyhedron shape, large and sufficient pseudopodia, abundant granules in the cytoplasm, narrow intercellular spaces and densely populated features. ( C ) LPS (0.5 μg/ml)‐treated pMAC s showed excessive activation: they presented with irregular, doublet or multiple shapes and ultimately underwent cell death, which was characterized by cell membrane blebbing, cell body atrophy and nuclear condensation or fragmentation.

    Article Snippet: The cells were harvested by centrifugation at 302 g for 10 min., re‐suspended in 5 ml of DMEM/F12 supplemented with 10% FBS and antibiotics, plated in 25 cm2 cell culture flasks (Corning), and maintained in a humidified atmosphere containing 5% CO2 at 37°C for 90 min.

    Techniques: Activation Assay

    Quantitative analysis of nine cytokines/enzymes secreted by pMAC s after exposure to TCM , DMEM /F12 or LPS (0.5 μg/ml) for 24 or 48 hrs. The mean and SD were calculated from nine values from three independent experiments. The bars that do not share a letter represent data that are significantly different ( P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: In vitro morphology, viability and cytokine secretion of uterine telocyte‐activated mouse peritoneal macrophages

    doi: 10.1111/jcmm.12711

    Figure Lengend Snippet: Quantitative analysis of nine cytokines/enzymes secreted by pMAC s after exposure to TCM , DMEM /F12 or LPS (0.5 μg/ml) for 24 or 48 hrs. The mean and SD were calculated from nine values from three independent experiments. The bars that do not share a letter represent data that are significantly different ( P

    Article Snippet: The cells were harvested by centrifugation at 302 g for 10 min., re‐suspended in 5 ml of DMEM/F12 supplemented with 10% FBS and antibiotics, plated in 25 cm2 cell culture flasks (Corning), and maintained in a humidified atmosphere containing 5% CO2 at 37°C for 90 min.

    Techniques:

    Cell viability of pMAC s after 48 hrs of co‐culture. As demonstrated by increased OD values, TCM and LPS significantly activated pMAC s compared to DMEM /F12. Nevertheless, no significant difference was observed between TCM and LPS ( P > 0.05), although a slightly higher value was obtained for TCM . * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: In vitro morphology, viability and cytokine secretion of uterine telocyte‐activated mouse peritoneal macrophages

    doi: 10.1111/jcmm.12711

    Figure Lengend Snippet: Cell viability of pMAC s after 48 hrs of co‐culture. As demonstrated by increased OD values, TCM and LPS significantly activated pMAC s compared to DMEM /F12. Nevertheless, no significant difference was observed between TCM and LPS ( P > 0.05), although a slightly higher value was obtained for TCM . * P

    Article Snippet: The cells were harvested by centrifugation at 302 g for 10 min., re‐suspended in 5 ml of DMEM/F12 supplemented with 10% FBS and antibiotics, plated in 25 cm2 cell culture flasks (Corning), and maintained in a humidified atmosphere containing 5% CO2 at 37°C for 90 min.

    Techniques: Co-Culture Assay

    NeuT oncogene transformation enhanced ALA-induced PpIX fluorescence ( A ) Fluorescence spectra of MCF10A vector and NeuT cell lysates and PpIX standard (25 ng/mL in DMSO). Both vector and NeuT cells were incubated with 1 mM ALA in serum free medium for 4 h and lysed. Fluorescence spectra of cell lysates and PpIX standard were obtained using 400 ± 2.5 nm excitation. ( B – D ) Flow cytometer analysis showing forward scatter (FSC) (B), basal cell fluorescence without ALA (C), and a dose-dependent fluorescence increase after ALA incubation (D) in MCF10A vector and NeuT cells. Cells were cultured in serum free DMEM/F12 medium with or without ALA for 4 h and cell fluorescence was measured with flow cytometry. ALA-induced fluorescence increase, calculated by subtracting basal cell fluorescence without ALA from cell fluorescence after incubation with different doses of ALA, was fit with the Michaelis-Menten enzyme kinetics. Data are presented as mean ± SD from at least 3 experiments. *** p

    Journal: Oncotarget

    Article Title: Her2 oncogene transformation enhances 5-aminolevulinic acid-mediated protoporphyrin IX production and photodynamic therapy response

    doi: 10.18632/oncotarget.11058

    Figure Lengend Snippet: NeuT oncogene transformation enhanced ALA-induced PpIX fluorescence ( A ) Fluorescence spectra of MCF10A vector and NeuT cell lysates and PpIX standard (25 ng/mL in DMSO). Both vector and NeuT cells were incubated with 1 mM ALA in serum free medium for 4 h and lysed. Fluorescence spectra of cell lysates and PpIX standard were obtained using 400 ± 2.5 nm excitation. ( B – D ) Flow cytometer analysis showing forward scatter (FSC) (B), basal cell fluorescence without ALA (C), and a dose-dependent fluorescence increase after ALA incubation (D) in MCF10A vector and NeuT cells. Cells were cultured in serum free DMEM/F12 medium with or without ALA for 4 h and cell fluorescence was measured with flow cytometry. ALA-induced fluorescence increase, calculated by subtracting basal cell fluorescence without ALA from cell fluorescence after incubation with different doses of ALA, was fit with the Michaelis-Menten enzyme kinetics. Data are presented as mean ± SD from at least 3 experiments. *** p

    Article Snippet: Cell culture and transfection MCF10A human breast epithelial cells were routinely maintained in complete DMEM/F12 (50/50) medium (Mediatech, Manassas, VA) supplemented with 5% horse serum (Atlanta Biologicals), insulin 10 ug/mL, epidermal growth factor (EGF) 20 ng/mL, cholera toxin 100 ng/mL, hydrocortisone 0.5 ug/mL and 1% of antibiotics and antimycotics solution (Mediatech) at 37° C in a humidified 5% CO2 incubator.

    Techniques: Transformation Assay, Fluorescence, Plasmid Preparation, Incubation, Flow Cytometry, Cytometry, Cell Culture

    Her2/NeuT oncogene expression transformed MCF10A human breast epithelial cells ( A ) Differential interference contrast (DIC) images (60×) show distinct differences in cell morphology between MCF10A vector control and NeuT-transformed cells. ( B ) Her2/NeuT oncogene transformation altered cell signaling. MCF10A vector and NeuT cells were cultured in complete DMEM/F12 medium, serum free medium with or without 1 mM ALA for 4 h and lysed with lysis buffer. Cell lysates were examined by Western blot for Her2/Neu signaling molecules, EMT and tight junction markers, and glycolytic enzyme PDK1.

    Journal: Oncotarget

    Article Title: Her2 oncogene transformation enhances 5-aminolevulinic acid-mediated protoporphyrin IX production and photodynamic therapy response

    doi: 10.18632/oncotarget.11058

    Figure Lengend Snippet: Her2/NeuT oncogene expression transformed MCF10A human breast epithelial cells ( A ) Differential interference contrast (DIC) images (60×) show distinct differences in cell morphology between MCF10A vector control and NeuT-transformed cells. ( B ) Her2/NeuT oncogene transformation altered cell signaling. MCF10A vector and NeuT cells were cultured in complete DMEM/F12 medium, serum free medium with or without 1 mM ALA for 4 h and lysed with lysis buffer. Cell lysates were examined by Western blot for Her2/Neu signaling molecules, EMT and tight junction markers, and glycolytic enzyme PDK1.

    Article Snippet: Cell culture and transfection MCF10A human breast epithelial cells were routinely maintained in complete DMEM/F12 (50/50) medium (Mediatech, Manassas, VA) supplemented with 5% horse serum (Atlanta Biologicals), insulin 10 ug/mL, epidermal growth factor (EGF) 20 ng/mL, cholera toxin 100 ng/mL, hydrocortisone 0.5 ug/mL and 1% of antibiotics and antimycotics solution (Mediatech) at 37° C in a humidified 5% CO2 incubator.

    Techniques: Expressing, Transformation Assay, Plasmid Preparation, Cell Culture, Lysis, Western Blot