dulbecco s modified eagle s medium  (Millipore)

Journal: Journal of Nanobiotechnology

Article Title: Enhanced detection with spectral imaging fluorescence microscopy reveals tissue- and cell-type-specific compartmentalization of surface-modified polystyrene nanoparticles

doi: 10.1186/s12951-016-0210-0

Figure Lengend Snippet: Physical-chemical characterization of polystyrene NPs. Intensity weighted size distribution of carboxylated ( a ) and PEGylated ( b ) polystyrene nanoparticles measured by dynamic light scattering. Representative TEM images of the particles are shown in the top right panels of each DLS plot, scale bars represent 400 nm. c , d Aggregation–agglomeration properties and corona thickening of polystyrene nanoparticles was measured by DLS as a change in hydrodynamic size. Particles were incubated in inorganic ( c ) or biological solutions ( d ) for 96 h, and the size distribution was monitored. Color codes of samples are shown in the figure. Data are presented as mean ± standard deviation (n = 3). Particles showed no aggregation in distilled water or in PBS during the 96-h assay period ( c ). In contrast a time-dependent, heavy particle aggregation was found in serum-free DMEM (d). Incubation of nanoparticles with 10 % FBS also evoked an immediate size increase, but prevented large-scale aggregation ( d ). SDS-PAGE analysis confirmed the adsorption of serum proteins to both PS-COOH and PS-PEG nanoparticles after 1 h incubation in 10 % serum containing MEM (e) and verified reduced protein adsorption of PEG-coated nanoparticles after 24 h incubation ( f )

Article Snippet: The suspensions were further diluted 1:10 with PBS (pH 7.4), DMEM cell culture medium (Sigma-Aldrich, St. Louis, MO, USA) or DMEM supplemented with 10 % fetal bovine serum (Invitrogen/Gibco, Carlsbad, CA, USA).

Techniques: Transmission Electron Microscopy, Incubation, Standard Deviation, SDS Page, Adsorption

Journal:

Article Title: Rgs5 Targeting Leads to Chronic Low Blood Pressure and a Lean Body Habitus ▿

doi: 10.1128/MCB.01889-07

Figure Lengend Snippet: Signaling assays. Aortic smooth muscle cells were isolated from Rgs5 +/+ and Rgs5 −/− mice and cultured in Dulbecco's modified Eagle's medium-F12 containing 20% fetal calf serum. Cells were serum starved for 24 h

Article Snippet: Cells in culture were maintained in Dulbecco's modified Eagle's medium-F12 containing 20% fetal calf serum and 2 mM glutamine with penicillin and streptomycin and were serum starved for 24 h prior to the signaling assays.

Techniques: Isolation, Mouse Assay, Cell Culture, Modification

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Quantification of MSCA-1 + cells under DMEM- and MC-XF culturing conditions by FACS analysis. Unseparated JPCs were plated in culture dishes in DMEM medium. For the first test runs, cells were maintained in DMEM medium whereas the second series underwent stepwise FCS reduction and convertion to MC-XF medium. At the same time points (day 2, 5 and 9 after conversion from DMEM to MC-XF culture conditions), cells were detached from the dishes and the percentages of MSCA-1 + cells were determined by FACS analysis. Significant higher amounts of MSCA-1 + cells were detected under MC-XF culturing conditions at day 2 and 5 (p

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: FACS

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Detection of mineral deposition by mineralizing JPCs (of passage 6). The upper panel of the figure illustrates the beginning of mineralization at day 12 of osteogenic induction (ob) under both media conditions. Note that precipitate formation originates from only a few cells of the DMEM-cultured monolayer. In contrast, the MC-XF-cultivated monolayer seemed to be purer due to the appearance of mineralization potential originating from almost every cell of the monolayer. 4× magnification. The lower panel of the figure illustrates representative fluorescent stainings of hydroxyapatite formation by OsteoImage in DMEM- and MC-XF-cultured monolayers (growing within coated flasks) at day 20 of osteogenesis. JPCs cultivated under DMEM media conditions showed a stronger mineralization potential. 10× magnification.

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Expression patterns of DMEM- and MC-XF-cultured JPCs (growing within coated flasks) by flow cytometric analyses. Representative histograms and the average percentages (±STD) of positive cells for CD29, CD45, CD73, CD90 and CD105 expression by unseparated JPCs are illustrated.

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: Expressing, Cell Culture, Flow Cytometry

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Detection of mineral deposition by non-mineralizing JPCs (of passage 6). Representative fluorescent staining of hydroxyapatite formation by OsteoImage in DMEM- and MC-XF-cultured monolayers (growing within coated flasks) at day 20 of osteogenesis. Non-mineralizing JPCs showed mineralization capacity only under MC-XF but not DMEM media conditions. 10× magnification. Co = untreated cell, ob = osteogenic induced cells.

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: Staining, Cell Culture

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Quantitative analysis of gene expression levels in DMEM- and MC-XF-cultured JPCs at day 5 and 10 of osteogenesis (of passage 6, n = 4). Induction indices (x-fold) and significance values of alkaline phosphatase, Runx-2, type I collagen (alpha1-chain) and osteoprotegerin in osteogenic induced in comparison to untreated cells under both culture conditions are illustrated.

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: Expressing, Cell Culture

Journal: PLoS ONE

Article Title: Selection of Osteoprogenitors from the Jaw Periosteum by a Specific Animal-Free Culture Medium

doi: 10.1371/journal.pone.0081674

Figure Lengend Snippet: Life-monitoring measurements of cell proliferation by unseparated JPCs using the x-CELLigence system (ACEA Biosciences). JPCs of passage 4 derived from two different donors were seeded into special E-plates in DMEM/F12/10%FCS culture medium. Two days later (44 hours - each tick of the scale corresponds to 11 hours), a gradual FCS reduction was performed in one-half of the test runs (green and dark green), whereas the other half of the wells was further cultivated in DMEM/F12/10%FCS (red and coral). Nine days (297 hours) after the initiation of FCS reduction, MC-XF culture medium was added to the cells. The proliferation curve progression of DMEM-cultured JPCs (from two representative patients) is highlighted in red and coral and that of MC-XF-cultivated cells is highlighted in dark green and green. The right panel of the figure shows the cell morphology of DMEM- and MC-XF-cultured JPCs. Note the reduced cell size under MC-XF culture conditions (on uncoated dishes) leading to the significant decrease of cell impedance immediately after the addition of the MC-XF culture medium.

Article Snippet: Differentiation experiments DMEM-cultured JPCs (4×104 cells per well in 6-well plates) were treated with osteogenic medium (ob - DMEM/F12 containing 10% FCS, 10 mM β-glycerophosphate, 100 µM L-ascorbic acid 2-phosphate and 4 µm dexamethasone, Sigma-Aldrich) for 30 days.

Techniques: Derivative Assay, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells

doi: 10.3389/fbioe.2019.00321

Figure Lengend Snippet: Chondrogenic potential of human IPFSCs after FN1-KO. Human IPFSCs were chondrogenically induced in a pellet culture system for 18 days. The effect of fibronectin on chondrogenic capacity of human IPFSCs was evaluated using gross observation of 18-day pellets, Alcian blue staining (Ab) for sulfated GAGs and immunohistochemical staining (IHC) for type II collagen (Col2) (A) . qPCR was used to evaluate expression of chondrogenic marker genes ( SOX9, ACAN, COL2A1 , and PRG4 ) and hypertrophic marker genes ( COL10A1 and MMP13 ) (B) . GAPDH was used as an endogenous control. Data are shown as bar charts. *indicates a significant difference compared to the corresponding copGFP group ( P

Article Snippet: After overnight incubation (day 0), pellets were grown in a serum-free chondrogenic induction medium [high-glucose Dulbecco's modified Eagle's medium (DMEM) with 40 μg/ml proline (MilliporeSigma), 100 nM dexamethasone (MilliporeSigma), 100 U/ml penicillin, 100 μg/ml streptomycin, 0.1 mM ascorbic acid-2-phosphate, and 1 × ITS™ Premix (BD Biosciences)] with the supplementation of 10 ng/ml transforming growth factor beta3 (TGFβ3; PeproTech, Rocky Hill, NJ) for up to 18 days.

Techniques: Staining, Immunohistochemistry, Real-time Polymerase Chain Reaction, Expressing, Marker

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells

doi: 10.3389/fbioe.2019.00321

Figure Lengend Snippet: Chondrogenic potential of human IPFSCs after expansion on dECMs deposited by FN1-KO cells. Passage 15 human IPFSCs in a pellet culture system were compared for chondrogenic capacity after expansion on dECMs deposited by Cas9-sgFN1a/b transduced cells (sgFN1a ECM and sgFN1b ECM, respectively) with those deposited by copGFP (copGFP ECM) and those grown on TCP (TCP) as controls including gross observation of 18-day pellets, Alcian blue staining (Ab) for sulfated GAGs, and IHC for type II collagen (Col2) (A) . qPCR was used to evaluate expression of chondrogenic marker genes ( SOX9, ACAN, COL2A1 , and PRG4 ) and hypertrophic marker genes ( COL10A1 and MMP13 ) (B) . GAPDH was used as an endogenous control. Data are shown as bar charts. *indicates a significant difference compared to the corresponding copGFP group ( P

Article Snippet: After overnight incubation (day 0), pellets were grown in a serum-free chondrogenic induction medium [high-glucose Dulbecco's modified Eagle's medium (DMEM) with 40 μg/ml proline (MilliporeSigma), 100 nM dexamethasone (MilliporeSigma), 100 U/ml penicillin, 100 μg/ml streptomycin, 0.1 mM ascorbic acid-2-phosphate, and 1 × ITS™ Premix (BD Biosciences)] with the supplementation of 10 ng/ml transforming growth factor beta3 (TGFβ3; PeproTech, Rocky Hill, NJ) for up to 18 days.

Techniques: Staining, Immunohistochemistry, Real-time Polymerase Chain Reaction, Expressing, Marker