“Two-cell embryos were incubated in an equilibration solution comprising with Ham's F-10 media with 20% human serum albumin (HAS) that supplemented with 7.5% ethylene glycol (Sigma-Aldrich, Germany) and 7.5% dimethyl sulphoxide (Sigma-Aldrich, Germany) for 5-15 min at room temperature (RT) condition. After then, they were placed into the vitrification solution containing Ham's F-10 medium supplemented with 15% ethylene glycol, 15% dimethyl sulphoxide and 0.5 M sucrose (Merck, Darmstadt, Germany) for 50-60 sec at RT. Eventually, the embryos placed on the tip of the Cryotop (Kitazato, Japan) for storage in liquid nitrogen. Warming of embryos was performed by placing the Cryotop in TS solution (1 M sucrose) for 50-60 sec at RT and then into a dilution solution (0.5 M sucrose) for 3 min. The warmed embryos were placed into a washing solution (Ham's F-10 and 20% HSA) four times" (19). They were divided into two groups. Half of the embryos were cultured to blastocysts for assessing their viability using Hoechst and PI staining. The viability rate was calculated based on the ratio of blue cells (as living cells) to the total cells. The rest of the two-cell embryos were collected for examining the expression of Zp3, E-cad, and Ctnnb1 genes using molecular techniques.
Dimethyl sulphoxide
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, Italy, India, China, Switzerland
Dimethyl sulphoxide (DMSO) is a colorless, odorless, and polar aprotic solvent. It is commonly used in laboratory settings as a solvent and cryoprotectant. DMSO has a high boiling point and is miscible with a wide range of organic and inorganic solvents.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (23 mentions)
Methanol, by Merck Group (20 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (15 mentions)
Triton x 100, by Merck Group (10 mentions)
Ethanol, by Merck Group (8 mentions)
Chloroform, by Merck Group (8 mentions)
Penicillin streptomycin, by Merck Group (7 mentions)
Cremophor, by Merck Group (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Acetonitrile, by Merck Group (7 mentions)
Actinomycin d, by Merck Group (6 mentions)
Bovine serum albumin (bsa), by Merck Group (6 mentions)
Mtt reagent, by Merck Group (6 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (6 mentions)
Sodium hydroxide (naoh), by Merck Group (6 mentions)
Quercetin, by Merck Group (6 mentions)
Fetal bovine serum (fbs), by Merck Group (6 mentions)
Acetone, by Merck Group (6 mentions)
Ethyl acetate, by Merck Group (6 mentions)
Trypsin edta, by Thermo Fisher Scientific (6 mentions)
232 protocols using dimethyl sulphoxide
1
Vitrification and Warming of Two-Cell Embryos
2
Antimicrobial Susceptibility Testing
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Chemicals used in the study included ciprofloxacin, dimethyl sulphoxide (DMSO), iodonitrotetrazolium (INT), and crystal violet were purchased from Sigma-Aldrich (Darmstadt, Germany). Tryptic soy broth (TSB) and tryptic soy agar (TSA) (22091) were also purchased from Sigma-Aldrich (Darmstadt, Germany).
3
DHEA Attenuates FeCl3-Induced Epilepsy in Rats
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A total of 18 rats were randomly assigned to three experimental groups. (1) Control: (n = 6) rats received an intracortical saline injection as FeCl3 solvent. (2) Epileptic: (n = 6) rats received an intracortical injection of FeCl3 solution as described above. (3) Epileptic + DHEA: (n = 6) rats received an intracortical injection of FeCl3 solution and DHEA treatment for 21 consecutive days.
After 20 days of FeCl3 injection, DHEA, solubilized in 0.1% dimethylsulphoxide (Sigma Aldrich, St. Louis, MO, USA), was injected intraperitoneally (30 mg/kg b. wt.) for the next 21 days (Figure 1 ). DHEA dose and duration used in this study are consistent with our laboratory’s earlier publications indicating its antiepileptic effect at 7, 14, and 21 days of treatment [8 (link),33 (link)]. As 21 days of DHEA administration showed a stronger antiepileptic effect, the same was chosen.
After 20 days of FeCl3 injection, DHEA, solubilized in 0.1% dimethylsulphoxide (Sigma Aldrich, St. Louis, MO, USA), was injected intraperitoneally (30 mg/kg b. wt.) for the next 21 days (
4
Preparation of Benzoic Acid and β-Cyclodextrin Solutions
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The following chemical reagents were used in this work:
Benzoic acid (C6H5COOH) (“chemically pure” grade), used without additional purification. The purity of reagents was declared by the manufacturer >99% by weight.
β-cyclodextrin (C42H70O35) from Sigma-Aldrich Saint (Louis, MO, USA) with a CD content of ≥99% was used without additional purification;
Dimethylsulfoxide (C2H6OS) was purified by distillation according to the method [75 ] before use. The DMSO content was 99.4 wt.%. The residual water content in the organic solvents used was taken into account when preparing the solutions.
Deuterated water and dimethylsulphoxide (atomic fraction of deuterium more than 99.9%) were purchased from Sigma-Aldrich USA. Ethyl alcohol (C2H5OH) (96% by vol.) was purified by distillation at the atmospheric pressure.
5
MTT Assay for Cell Viability
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In each well of 96‐well plates, 3 × 103 of tumour cells were treated by 5 mg mL–1 of 2‐(4,5‐dimethyltriazol‐2‐yl)−2,5‐diphenyl tetrazolium bromide (MTT, Sigma). After 4 h of incubation at 37°C, cell supernatants were removed. MTT crystals were dissolved in 150 µL of dimethylsulphoxide (Sigma), while absorbance was measured at 570 nm.53
6
MTT Assay for Cell Viability
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The rate of metabolically active cells was evaluated using MTT assay, according to the manufacturer’s protocol. Cells were seeded at a density of 8x103 cells/well in 96-well plates (Corning Inc., Corning, NY) and cultured with different treatments for defined duration at 37°C in a humidified CO2 incubator. Cells were incubated with solution of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; Sigma, St. Louis, MO) for 4 h. Supernatant was removed and dimethyl sulphoxide (Sigma, St. Louis, MO) was added to dissolve the purple formazan crystals. The absorbance was measured at 570 nm with a microplate reader (Tecan microplate reader, Infinite M200 Pro). Cell viability was estimated as a percentage of the value of the untreated control cells.
7
Investigating circFOXM1 and FOXM1 mRNA Expression
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To block transcription, 2 mg/mL actinomycin D or dimethylsulphoxide (Sigma-Aldrich, St. Louis, MO, USA) as a negative control was added into the cell culture medium. For RNase R treatment, total RNA (2 μg) was incubated for 30 min at 37°C, with or without 3 U/μg of RNase R (Epicenter Technologies, Madison, WI, USA). After treatment with actinomycin D and RNase R, qRT-PCR was performed to determine the expression levels of circFOXM1 and FOXM1 mRNA.
8
MTT Assay for Cell Proliferation and Drug Resistance
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MTT assay was first conducted to investigate cell proliferation in CPA4 silencing or overexpressing PC cells at different time points (one to five days) combining with or without LY294002 treatment. CPA4siRNA transfected PANC-1 cells and CPA4-FLAG transfected Capan-2 cells (with or without 10 µm LY294002 treatment three times) were seeded into 96-well plates at the density of 5000 viable cells per well and incubated for one to five days. At the end of each time point, 15 µL of MTT (5 mg/mL in PBS, Sigma) was added for four hours and the medium was then incubated with 100 µL of dimethyl sulphoxide (Sigma) was added to each well for 20 min. Drug resistance assays were done in a similar way with various concentrations of gemcitabine (GEM, Abcam) as shown in results section. Per experiment group at a wavelength of 570 nm in an ELISA 96-well microtiter plate reader (BIORAD680, USA). Experiments were performed in triplicate, and data were presented as a percentage of treated cells compared with control cells.
9
Detailed Protocol of Cell Culture Reagents
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1-Aminobenzotriazole, ammonium chloride, ciprofoxacin, clarithromycin, dimethyl sulphoxide, Imipramine, lactate dehydrogenase activity assay kit, monensin sodium salt, nigericin sodium salt, trypan blue 0.4% and verapamil hydrochloride were all from Sigma Aldrich Ltd., Dorset, UK. Budesonide, ipratropium bromide, fenoterol, formoterol, terbutaline and tiotropium bromide were all supplied by GlaxoSmithKline, UK. Chloroform and formaldehyde 37–41% were from Fisher Scientific, Loughborough, UK. Further chemicals include diazepam (Tocris Bioscience, Bristol, UK), methanol (VWR, UK), midazolam (Hoffman La Roche, Switzerland), Pierce BCA protein assay kit (Thermo Scientific, Loughborough, UK) and Lysotracker® Red DND-99 (Life Technologies, Paisley, UK). Reagents include bovine serum albumin and penicillin-streptomycin (Sigma Aldrich Ltd., Dorset, UK), collagen Type I rat tail (BD Biosciences, Oxford, UK), Dulbecco’s phosphate buffered saline and heat-inactivated foetal bovine serum (Life Techonolgies, Paisley, UK) and Kaighn’s modification of Ham’s F12 (Ham’s F12K) medium (American Tissue Culture Collection (ATCC), Mannasas, VA, USA).
10
Pericyte Cell Culture and Characterization
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Heat inactivated FBS (fetal bovine serum), antibiotic–antimycotic, Dulbecco’s phosphate buffered saline (DPBS), high glucose (hg) DMEM, M199, Endothelial Cell Growth Medium (ECM) and Geltrex™ LDEV-Free Reduced Growth Factor Basement Membrane Matrix and NuPage 4–12% bis-Tris Gel, were purchased from Gibco-Life Technologies (Carlsbad CA, USA). Trypsin–EDTA solution1X, Dimethyl sulphoxide (DMSO), lipopolysaccharide (LPS) (E. coli 055:B5), Protein Assay Kit TP0300, In Vitro Toxicology Assay Kit and Cell Growth Determination Kit MTT based were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pericyte Growth Medium was purchased from Promocell (Heidelberg, Germany). NucleoSpin RNA kit was purchased from Macherey-Nagel GmbH & Co. KG (Düren, Germany) RT2 strand kit, RT2 Sybr green fluor qPCR master mix were from Qiagen (Hilden, Germany). Porcine Cytokine/Chemokine Magnetic Bead Panel kit, Milliplex Map Kit EMD was purchased by Millipore Corporation (Billerica, MA, USA). Super Signal West Pico Chemiluminescent Substrate was from Pierce Biotechnology, Inc. (Rockford, IL, USA).
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