Confluent monolayers of HUVEC or BAEC were trypsinized. Cells were suspended in corresponding culture medium containing 20% methocel, seeded into nonadhesive 75-cm2 bacteriological dishes (Greiner, Frickenhausen, Germany), and cultured at 37°C (5% CO2, 100% humidity). Under these conditions suspended EC aggregate spontaneously within 4 h to form cellular aggregates of varying size and cell number. The methocel used for these experiments was diluted from a stock solution that was generated by dissolving 6 g of carboxymethylcellulose in 500 ml of medium (DME or ECGM basal medium). After centrifugation the clear, gel-like supernatant was used for experiments. Methocel prevents adhesion of cells and acts as an inert viscosity modulating substance. Variation of the methocel concentration during spheroid formation was, thus, used to control the average size of the spheroids. These multicellular spheroids were designated as random spheroids and used for all experiments that employed larger populations of cells. To generate endothelial cell spheroids of defined size and cell number, a specific number of cells (varying between 500 and 3,000 cells per spheroid, depending on the experiment) was suspended in culture medium and seeded in nonadherent round-bottom 96-well plates (Greiner, Frickenhausen, Germany). Under these conditions all suspended cell contribute to the formation of a single endothelial cell spheroid. These spheroids, designated as standard spheroids, were harvested within 24 h and used for the corresponding experiments.
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Methocel
Methocel
Methocel, also known as methylcellulose, is a water-soluble cellulose ether used in a variety of pharmaceutical and industrial applications.
It is commonly employed as a thickening, suspending, and stabilizing agent in topical, oral, and injectable formulations.
Methocel helps to control viscosity, improve texture, and enhance the bioavailability of active ingredients.
It is also utilized in food products, cosmetics, paints, and other materials to modify rheological properties.
Researchers can optimize their Methocel experiments using the PubCompare.ai platform, which uses AI-powered intelligent comparisons to help locate the best protocols from literature, preprints, and patents, ensuring accurate and reproducible results and taking the guesswork out of Methocel research.
It is commonly employed as a thickening, suspending, and stabilizing agent in topical, oral, and injectable formulations.
Methocel helps to control viscosity, improve texture, and enhance the bioavailability of active ingredients.
It is also utilized in food products, cosmetics, paints, and other materials to modify rheological properties.
Researchers can optimize their Methocel experiments using the PubCompare.ai platform, which uses AI-powered intelligent comparisons to help locate the best protocols from literature, preprints, and patents, ensuring accurate and reproducible results and taking the guesswork out of Methocel research.
Most cited protocols related to «Methocel»
Carboxymethylcellulose
Cell Adhesion
Cells
Centrifugation
Culture Media
Endothelial Cells
Endothelium
Humidity
Hyperostosis, Diffuse Idiopathic Skeletal
Methocel
Population Group
SERPINA3 protein, human
Spheroids, Cellular
Viscosity
The 3T3-L1 cell (#EC86052701-G0, KAK) is a universally used cell line for lipid studies. The 3T3-L1 preadipocytes were grown until confluence at 37 °C in HG- DMEM containing 8 mg/L d-biotin, 4 mg/L calcium pantothenate, 100 U/mL penicillin, 100 μg/mL streptomycin (b.p. HG-DMEM), and 10% CS.
The 3T3-L1 organoids were generated by a hanging droplet spheroid three dimension (3D) culture system as described recently28 (link). Briefly, to facilitate stable morphology, methylcellulose (Methocel A4M) was added to the growth medium. Prior to seeding the hanging drop culture plate (# HDP1385, Sigma-Aldrich), cells were cultured in 100 mm or 150 mm dishes until reaching approximately 90% confluence. After washing with a phosphate buffered saline (PBS), cells were detached using 0.25% Trypsin/EDTA and resuspended in growth medium. After centrifugation for 5 min at 300 g, the cell pellet was re-suspended in a growth medium containing 0.25% w/v Methocel A4M. Volume was adjusted so that 20,000 cells were contained in the 28 μL solution, and 28 μL drops were placed into each well of the drop culture plate (defined as 3D/Day 0). An organoid medium (i.e. growth medium with 0.25% w/v Methocel A4M) was used throughout the duration of the spheroid culture. On every following day, 14 μL of the culture medium was removed and a fresh 14 μL culture medium was added to each well.
The 3T3-L1 organoids were generated by a hanging droplet spheroid three dimension (3D) culture system as described recently28 (link). Briefly, to facilitate stable morphology, methylcellulose (Methocel A4M) was added to the growth medium. Prior to seeding the hanging drop culture plate (# HDP1385, Sigma-Aldrich), cells were cultured in 100 mm or 150 mm dishes until reaching approximately 90% confluence. After washing with a phosphate buffered saline (PBS), cells were detached using 0.25% Trypsin/EDTA and resuspended in growth medium. After centrifugation for 5 min at 300 g, the cell pellet was re-suspended in a growth medium containing 0.25% w/v Methocel A4M. Volume was adjusted so that 20,000 cells were contained in the 28 μL solution, and 28 μL drops were placed into each well of the drop culture plate (defined as 3D/Day 0). An organoid medium (i.e. growth medium with 0.25% w/v Methocel A4M) was used throughout the duration of the spheroid culture. On every following day, 14 μL of the culture medium was removed and a fresh 14 μL culture medium was added to each well.
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3T3-L1 Cells
Biotin
Cell Lines
Cells
Centrifugation
Culture Media
Edetic Acid
Hyperostosis, Diffuse Idiopathic Skeletal
Lipid A
Methocel
Methocel E
Methylcellulose
Organoids
Pantothenate, Calcium
Penicillins
Phosphates
Saline Solution
Streptomycin
Trypsin
HOF cells prepared as above were cultured in 100-mm or 150-mm dishes until reaching 90% to 100% confluence, washed with PBS twice, detached by 0.25% trypsin-EDTA, resuspended with a growth medium, and centrifuged at 300×g for 5 minutes. 3D organoid preparations were carried out on hanging drop culture plates (HDP1385; Sigma-Aldrich, St. Louis, MO, USA) as described recently.32 (link) Briefly, the cell pellet obtained as above was resuspended in a growth medium containing 0.25% METHOCEL (DuPont Nutrition & Biosciences, Copenhagen, Denmark) to stimulate morphological stability. Approximately 20,000 cells in a 28-µL drop were placed in each well of a hanging drop culture plate (day 0). Thereafter, 14 µL of the culture medium was removed and a fresh 14 µL of the culture medium was added to each well every day until day 12 for HOFs. During the course of the 3D culture, HOFs grew into a sphenoid-shaped organoid. Adipogenic differentiation of HOFs was induced by a cocktail containing 250-nM dexamethasone, 10-nM T3, 10 µg/ml insulin, and 10-mM troglitazone from day 1 through day 5. This was followed by the use of a cocktail containing 10 µg/ml insulin and 10-mM troglitazone from day 6 through day 11 for HOFs in the presence or absence of 100-µM BIM or 100-µM prostaglandin F2α (PGF2α). 3D cell organoids of HOFs at day 12 were collected for RNA preparation or immunostaining. As a control, 3D culture cells were treated as above in the growth medium containing 0.1% dimethyl sulfoxide without the adipogenic stimulation.
Adipogenesis
Cell Culture Techniques
Cells
Culture Media
Dexamethasone
Dinoprost
Edetic Acid
Hyperostosis, Diffuse Idiopathic Skeletal
Insulin
Methocel
Organoids
Sphenoid Bone
Sulfoxide, Dimethyl
Troglitazone
Trypsin
The animals were dark adapted overnight before the ERG recordings, and their manipulation was done under dim red light (λ>600 nm). The rats were anaesthetized and bilateral pupil mydriasis was induced by applying a topical drop of 1% tropicamide (Colircusi tropicamida 1%®; Alcon-Cusí, S.A., El Masnou, Barcelona, Spain) to both eyes. The light stimulation device used was a Ganzfeld dome, which ensures a homogeneous illumination anywhere in the retina, with multiple reflections of the light generated by light emitting diodes (LED), which provided a wide range of light intensities. For high intensity illuminations, a single LED placed close (1 mm) to the eye was used. Light intensity was calibrated by a dual-biosignal generator device specifically adapted for ERG responses. The recording system comprised Burian-Allen bipolar electrodes (Hansen Labs, Coralville, IA) with a corneal contact shape; a drop of methylcellulose, 2% (Methocel 2%®; Novartis Laboratories CIBA Vision, Annonay, France) was placed between the eye and the electrode to maximize conductivity of the generated response. The reference electrode was placed in the mouth and the ground electrode in the tail. Electrical signals generated in the retina were amplified (x1000) and filtered (band pass from 1 Hz to 1000 Hz) by a commercial amplifier (Digitimer Ltd, Letchworth Garden City, UK). The recorded signals were digitized (Power Lab; ADInstruments Pty. Ltd., Chalgrove, UK) and displayed on a PC computer. Bilateral ERG recording was performed simultaneously from both eyes. Light stimuli were calibrated before each experiment, and the calibration protocol was set to assure the same recording parameters for both eyes. The ERG responses were recorded by stimulating the retina with light intensities ranging between10−6 and 10−4 cd·s·m−2 for the scotopic threshold response (STR), 10−4 and 10−2 cd·s·m−2 for the rod response, and 10−2 and 102 cd·s·m−2 for the mixed (rod and cone) response. For each light intensity level, a series of ERG responses were averaged (from 40 ERG responses for the dimmest stimulus intensities to 5 for the brightest stimulus), and the interval between light flashes was adjusted to ensure a timing that allowed response recovery (from 5 s for the dimmest stimulus intensities to 60 s for the brightest stimulus). At the end of each session the animals were treated with topical tobramicine (Tobrex®; Alcon-Cusí, S.A.) in both eyes. The analysis of the different recordings was performed with the normalization criteria established for the International Society for Clinical Electrophysiology of Vision (ISCEV) for the measures of the amplitude and implicit time of the different waves studied.
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Animals
Colircusi Tropicamida
Cornea
Electric Conductivity
Electricity
Eye
Light
Lighting
Medical Devices
Methocel
Methylcellulose
Mydriasis
Oral Cavity
Photic Stimulation
Pupil
Rattus
Reflex
Retina
Retinal Cone
Tail
Tropicamide
Vision
The 3T3-L1 preadipocytes (#EC86052701-G0, KAK), a cell line that is universally used in lipid studies, were cultured in 2D culture dishes at 37 °C in a grown medium; high glucose Dulbecco’s modified Eagle medium (HG-DMEM, FUJIFILM, Osaka, Japan) supplemented with 8 mg/L d-biotin, 4 mg/L calcium pantothenate, 100 U/mL penicillin, 100 μg/mL streptomycin (b.p. HG-DMEM), 10% CS and methylcellulose (Methocel A4M, Sigma-Aldrich, St. Louis, MO, USA) as a morphology stabilizer.
To generate 3D spheroids under culture conditions identical to those used for the 2D cell culture except culture plates, a hanging droplet spheroid 3D culture system, described recently [6 (link),14 (link)], was used. Briefly, 3T3-L1 cells were cultured in 100 mm or 150 mm dishes as described above. After reaching approximately 90% confluence, the cells were detached by treatment with 0.25% trypsin/EDTA after washing with phosphate-buffered saline (PBS) and then resuspended in the growth medium. These cell suspensions were divided into conventional 2D cultures and 3D spheroid cultures. The 2D-cultured 3T3-L1 cells were again maintained 100 mm 2D culture dishes until Day 7, with medium changes daily. Alternatively, a cell suspension containing approximately 20,000 cells in a 28 μL culture medium was then placed into each well of the 3D drop culture plate (# HDP1385, Sigma-Aldrich, St. Louis, MO, USA). This timing was defined as 3D/Day 0, and 14 μL of the culture medium was then replaced with 14 μL of fresh culture medium in each well daily until Day 7 [7 (link),8 (link),10 (link)]. On Day 7, both 2D- and 3D-cultured 3T3-L1 cells were each collected and further processed for use in the RNA-sequencing analysis, as described below.
To generate 3D spheroids under culture conditions identical to those used for the 2D cell culture except culture plates, a hanging droplet spheroid 3D culture system, described recently [6 (link),14 (link)], was used. Briefly, 3T3-L1 cells were cultured in 100 mm or 150 mm dishes as described above. After reaching approximately 90% confluence, the cells were detached by treatment with 0.25% trypsin/EDTA after washing with phosphate-buffered saline (PBS) and then resuspended in the growth medium. These cell suspensions were divided into conventional 2D cultures and 3D spheroid cultures. The 2D-cultured 3T3-L1 cells were again maintained 100 mm 2D culture dishes until Day 7, with medium changes daily. Alternatively, a cell suspension containing approximately 20,000 cells in a 28 μL culture medium was then placed into each well of the 3D drop culture plate (# HDP1385, Sigma-Aldrich, St. Louis, MO, USA). This timing was defined as 3D/Day 0, and 14 μL of the culture medium was then replaced with 14 μL of fresh culture medium in each well daily until Day 7 [7 (link),8 (link),10 (link)]. On Day 7, both 2D- and 3D-cultured 3T3-L1 cells were each collected and further processed for use in the RNA-sequencing analysis, as described below.
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3T3-L1 Cells
Biotin
Cell Culture Techniques
Cell Lines
Cells
Culture Media
Eagle
Edetic Acid
Glucose
Hyperostosis, Diffuse Idiopathic Skeletal
Lipids
Methocel
Methylcellulose
Pantothenate, Calcium
Penicillins
Phosphates
Saline Solution
Sequence Analysis
Streptomycin
Trypsin
Most recents protocols related to «Methocel»
The materials used in this
experiment include locust bean gum (Viscogum, Cargill, France), citric
acid monohydrate (Brand KgaA, Darmstadt, Germany), hydrochloric acid
(Sigma-Aldrich Chemie, GmbH), distilled water (sterilized water for
injection, PT. Otsuka Indonesia), acetone (Cawan Anugerah Chemika,
Indonesia), hydroxypropyl methylcellulose (Methocel K4M CR Premium
USP/EP, Colorcon, Singapore), lactose monohydrate (Leprino Foods,
UDM), ketoprofen (PT Kalbe Farma Tbk, Indonesia), potassium dihydrogen
phosphate (KGaA Darmstadt Germany Brand), and sodium hydroxide (KGaA
Darmstadt Germany Brand).
experiment include locust bean gum (Viscogum, Cargill, France), citric
acid monohydrate (Brand KgaA, Darmstadt, Germany), hydrochloric acid
(Sigma-Aldrich Chemie, GmbH), distilled water (sterilized water for
injection, PT. Otsuka Indonesia), acetone (Cawan Anugerah Chemika,
Indonesia), hydroxypropyl methylcellulose (Methocel K4M CR Premium
USP/EP, Colorcon, Singapore), lactose monohydrate (Leprino Foods,
UDM), ketoprofen (PT Kalbe Farma Tbk, Indonesia), potassium dihydrogen
phosphate (KGaA Darmstadt Germany Brand), and sodium hydroxide (KGaA
Darmstadt Germany Brand).
Acetone
Food
Hydrochloric acid
Hypromellose
Ketoprofen
Lactose
locust bean gum
Methocel
potassium phosphate, monobasic
Sodium Hydroxide
For angiogenesis assays, the spheroids were embedded in collagen gels composed of 8 parts of collagen, 1 part of M199 10×, and 1 part of NaOH [32 (link)].
Spheroids from each 96-well plate were collected with a 1000 μL pipette tip, transferred into a 50 mL tube, and then centrifuged at 300× g for 3 min. The supernatant was discarded and the spheroids were gently re-suspended in 1 mL medium M199 + 20% FCS + 0.5% methylcellulose (6 mL methocel solution + 600 μL FCS + 7.8 mL M199 (including 20% FCS)). The same volume of collagen gel (1:1) was added and 1 mL of this mixture was plated in one well of a 24-well plate. Incubation took place at 37 °C. Images were taken via fluorescence microscopy after 1 h and 24 h. Quantification of sprout length was performed with Image J [39 (link)] using Fiji distribution and the newest version available. The plugin “angiogenesis” was applied and within this plugin we used the plugin “sprout analysis” according to Eglinger et al. [40 (link)].
Spheroids from each 96-well plate were collected with a 1000 μL pipette tip, transferred into a 50 mL tube, and then centrifuged at 300× g for 3 min. The supernatant was discarded and the spheroids were gently re-suspended in 1 mL medium M199 + 20% FCS + 0.5% methylcellulose (6 mL methocel solution + 600 μL FCS + 7.8 mL M199 (including 20% FCS)). The same volume of collagen gel (1:1) was added and 1 mL of this mixture was plated in one well of a 24-well plate. Incubation took place at 37 °C. Images were taken via fluorescence microscopy after 1 h and 24 h. Quantification of sprout length was performed with Image J [39 (link)] using Fiji distribution and the newest version available. The plugin “angiogenesis” was applied and within this plugin we used the plugin “sprout analysis” according to Eglinger et al. [40 (link)].
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angiogen
Biological Assay
Collagen
Methocel
Methylcellulose
Microscopy, Fluorescence
Spheroid preparation was performed according to Augustin and Korff 1998 [31 (link)]. Six g methocel was dissolved in 500 mL M199 medium (supplemented with 1% L-glut, 1% PS and 10% FCS). The combined solution was stirred overnight at 4 °C and centrifuged at 3500× g for 3 h. Meanwhile, the cells were labelled with Cell Tracker™ according to the manufacturer’s instructions (hOBs with Cell Tracker™ Green and HUVECs with Cell Tracker™ Red). One part of the methocel solution (2.4 mL/96-well plate) was diluted with four parts medium (9.6 mL/96-well plate) and the cell suspension (for 1 × 96-well: 6 × 104 cells per mono-culture or 3 × 104 of each cell type per co-culture) was added. After gently mixing the cells, they were seeded in a 96-well plate with U-bottom (100 μL/well corresponding to 500 cells/well) and incubated overnight. On the next day, the spheroids had formed.
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5-chloromethylfluorescein diacetate
Cell Culture Techniques
Cells
Coculture Techniques
Human Umbilical Vein Endothelial Cells
Methocel
SLC2A1 protein, human
Levocetirizine dihydrochloride was a kind gift from Egis Pharmaceuticals PLC (Budapest, Hungary). Beta-cyclodextrin (β-CD), randomly methylated-beta-cyclodextrin (RAMEB), sulfobutylated-beta-cyclodextrin sodium salt (SBECD) and (hydroxypropyl)-beta-cyclodextrin (HPBCD) were obtained from Cyclolab Ltd. (Budapest, Hungary). Hydroxypropyl methylcellulose (Methocel E5, HPMC) and polyvinyl-pyrrolidone (Mw = 40,000) were purchased from Colorcon Ltd. (Budapest, Hungary) and Alfa Aesar (Kandel, Germany), respectively. Polyvinyl alcohol (Mw = 30,000–70,000) was a product of Sigma-Aldrich Inc. (Budapest, Hungary). Sodium hydroxide was delivered by Reanal Ltd. (Budapest, Hungary). Isopropyl myristate (IPM) was obtained from Sigma Aldrich (Budapest, Hungary). Phosphate buffer at pH = 7.4 was used as a reference. The PBS was composed of NaCl, KCl, CaCl2, Na2HPO4 and KH2PO4, and the SNES consisted of NaCl, KCl and anhydrous CaCl2 in deionized water, used at pH = 6.0 ± 0.1 with HCl. All these materials were purchased from Sigma-Aldrich (Budapest, Hungary).
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2-Hydroxypropyl-beta-cyclodextrin
betadex
Buffers
Hypromellose
isopropyl myristate
levocetirizine dihydrochloride
Methocel
Necrotizing encephalopathy, infantile subacute, of Leigh
Pharmaceutical Preparations
Phosphates
Polyvinyl Alcohol
Povidone
Sodium
Sodium Chloride
Sodium Hydroxide
We purchased micronized griseofulvin (BP/EP grade, BCS Class II, GF) from Letco Medical (Decatur, AL, USA). The aqueous solubility of GF is 14.2 mg/L at 37 °C [40 (link)]. Its melting point Tm and glass transition temperature Tg are 220 °C and 89 °C, respectively [41 (link)]. Three polymers and one surfactant were used in the formulations. BASF (Tarrytown, NY) donated Soluplus® (Sol), which is a graft copolymer made of polyvinyl caprolactam–polyvinyl acetate–polyethylene glycol, and its Tg is 73 °C [42 (link)]. Kollidon VA64 (VA64) was also donated by BASF, which is a vinylpyrrolidone-vinyl acetate copolymer with a Tg of 102 °C [43 ]. Dow Chemicals (Midland, MI, USA) donated hydroxypropyl methyl cellulose (HPMC, Methocel-E3 grade), which is a nonionic cellulosic polymer with a Tg of 174 °C [44 (link)]. We purchased sodium dodecyl sulfate (ACS grade, SDS) from GFS chemicals (Columbus, OH, USA), which is an anionic surfactant used to enhance the wettability of the ASD particles in the dissolution tests. We purchased acetone (ACS reagent, ≥99.5%) and ethanol (reagent alcohol, ≥95%) from BDH Analytical chemicals (Radnor, PA, USA) and used them as solvents to prepare drug–polymer solutions. We purchased the milling media, yttrium zirconia beads (Zirmil Y), from Saint Gobain ZirPro (Mountainside, NJ, USA), which had a median size of 430 μm.
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Acetone
Caprolactam
Ethanol
Grafts
Griseofulvin
Hypromellose
Kollidon VA64
Methocel
Pharmaceutical Solutions
Polyethylene Glycols
Polymers
polyvinyl acetate
soluplus
Solvents
Sulfate, Sodium Dodecyl
Surface-Active Agents
vinylacetate-vinylpyrrolidone copolymer
Vitrification
Yttrium
zirconium oxide
Top products related to «Methocel»
Sourced in United States, Germany
Methocel is a cellulose-based polymer material used in various laboratory applications. It serves as a thickening, suspending, and binding agent in pharmaceutical, food, and other industrial formulations.
Sourced in United States, Germany, United Kingdom, India, Italy, France, Sao Tome and Principe, Spain, Poland, China, Belgium, Brazil, Switzerland, Canada, Australia, Macao, Ireland, Chile, Pakistan, Japan, Denmark, Malaysia, Indonesia, Israel, Saudi Arabia, Thailand, Bangladesh, Croatia, Mexico, Portugal, Austria, Puerto Rico, Czechia
Tween 80 is a non-ionic surfactant and emulsifier. It is a viscous, yellow liquid that is commonly used in laboratory settings to solubilize and stabilize various compounds and formulations.
Sourced in United States, Germany, United Kingdom, Japan, Belgium, Switzerland, Italy, Australia, Canada, China, Sao Tome and Principe, Poland, Israel, France, Spain, Austria
Methylcellulose is a water-soluble, synthetic polymer derived from cellulose. It is a white, odorless, and tasteless powder that is commonly used as a thickening, stabilizing, and emulsifying agent in various industries, including pharmaceutical, food, and personal care products.
Sourced in Cameroon, United States
Methocel is a cellulose-based polymer product manufactured by Dow. It is a versatile material used in a variety of applications, including as a binder, thickener, and stabilizer in laboratory settings.
Sourced in United States
Methocel A4M is a cellulose-based excipient used in pharmaceutical formulations. It functions as a binder, thickener, and viscosity-modifying agent. The specific details of its core function and intended use are not provided, as maintaining an unbiased and factual approach without interpretation or extrapolation is required.
Methocel E4M is a cellulose-based product manufactured by Dow. It functions as a thickening, suspending, and film-forming agent for various applications.
Sourced in United States
METHOCEL A15LV is a hydroxypropyl methylcellulose product manufactured by Merck Group. It is a white to off-white, free-flowing powder used as an excipient in pharmaceutical and food applications.
Sourced in United States
Methocel 60 HG is a pharmaceutical grade cellulose derivative product manufactured by Merck Group. It is a fine, white to off-white, free-flowing powder. Methocel 60 HG is used as a binder, thickener, and stabilizer in various pharmaceutical and personal care formulations.
Sourced in United States
METHOCEL E4M Premium is a type of methylcellulose, a water-soluble cellulose ether. It is used as a thickening, suspending, and binding agent in various applications.
Sourced in United Kingdom
Methocel K4M is a cellulose-based polymer that functions as a binder, thickener, and film-forming agent in pharmaceutical and food applications. It is a white to off-white, granular powder that is soluble in water and other polar solvents. The specific viscosity of Methocel K4M can be adjusted through manufacturing parameters.
More about "Methocel"
Methylcellulose, Methocel A4M, Methocel E4M, METHOCEL A15LV, Methocel 60 HG, METHOCEL E4M Premium, Tween 80