Rat astrocytic C6 glioma cells and human embryonic kidney (HEK)-293 cells were purchased from RIKEN Cell Bank (Saitama, Japan). Mouse embryonal carcinoma P19 cells were obtained from ATCC (Manassas, VA, USA). Mouse microglial BV-2 cells are a generous gift from Dr. Eui-Ju Choi (Korea University, Seoul, Korea) [28 (link)]. Poly-L-lysine, all-trans retinoic acid (ATRA), Hoechst33342, propidium iodide (PI), A23187, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), 2-mercaptopyridine N-oxide sodium (pyrithione) and N,N,N’,N’-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) were purchased from Sigma-Aldrich fine Chemicals (St. Louis, MO, USA). Acetoxymethyl esters of Fluo-3, Rhod-2 and FluoZin-3 were provided by Molecular Probes (Eugene, OR, USA). Both EGTA and BAPTA-AM were supplied by Dojindo (Kumamoto, Japan). Dulbecco’s Modified Eagle Medium (DMEM) and alpha minimal essential medium (αMEM) were provided by Wako (Osaka, Japan). EDTA was purchased from Nacalai Tesque (Kyoto, Japan). Other chemicals used were all of the highest purity commercially available.
Dulbecco s modified eagle medium dmem
Manufactured by Fujifilm
Sourced in Japan, United States
Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium commonly used in research laboratories. It provides a nutrient-rich environment for the growth and maintenance of various cell types. DMEM contains a balanced salt solution, amino acids, vitamins, and other essential components required for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Wst 8 reagent, by Fujifilm (2 mentions)
Mouse il 6 uncoated elisa kit, by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Recombinant bovine hmgb1, by Chondrex (2 mentions)
Penicillin, by Fujifilm (2 mentions)
Streptomycin, by Fujifilm (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Biowest (2 mentions)
Hek293 cells, by RIKEN Cell Bank (1 mentions)
N n n n tetrakis 2 pyridylmethyl ethylenediamine tpen, by Merck Group (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Dojindo Laboratories (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Nacalai Tesque (1 mentions)
Bapta am, by Dojindo Laboratories (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Fluozin 3, by Thermo Fisher Scientific (1 mentions)
Rhod 2, by Thermo Fisher Scientific (1 mentions)
26 protocols using dulbecco s modified eagle medium dmem
1
Investigating Cell Line Responses to Ion Modulators
2
MIA PaCa-2 cell culture protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MIA PaCa-2 cells were grown in high glucose Dulbecco’s modified Eagle medium (DMEM; FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS; Equitech-Bio Inc., Kerrville, TX, USA) under standard cell culture conditions (humidified atmosphere, 5% CO2, 37 °C). The cell media was then replaced with low-glucose DMEM supplemented with 10% FBS and cells at a density of 0.8 × 105 cells/mL were seeded in culture dishes and plates (TrueLine, Pittston, PA, USA), and cultured until use in experiments (excluding experiments under high-glucose conditions).
3
In vitro Syncytialization of Cytotrophoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HTR-8/SVneo cells were cultured on 24-well plates with Dulbecco’s modified Eagle medium (DMEM, Wako) with 10% fetal bovine serum (FBS) and antibiotic-antimycotic solution at 37 °C under 5% CO2. Human villous cytotrophoblast cells were isolated from normal term placentas, and in-vitro syncytialization of isolated cytotrophoblasts were conducted as previously reported11 (link). Briefly, only HLA-ABC negative cells were collected using a Mini MACS TM separator (Miltenyi Biotec). Purified cytotrophoblasts were diluted in Iscove’s Modified Dulbecco’s Medium (IMDM) (GE Healthcare) supplemented with 10% fetal bovine serum (FBS), 1% antibiotic-antimycotic (Thermo Fisher Scientific), 200 mM L-glutamine and 10 ng/ml epidermal growth factor (EGF). After seeding the isolated cytotrophoblasts, spontaneous cell fusion occurred simultaneous with syncytialization in vivo. After 96 hours in culture, the culture wells were covered by a monolayer of syncytialized trophoblast, then treated with 100 pg/ml recombinant human TNFSF14/LIGHT (R&D) for 18 hours.
4
Caco-2 Interleukin-8 Response to Flagellin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Caco-2 cells, originating from the human colon, were maintained in Dulbecco’s modified Eagle Medium (DMEM, Wako) supplemented with 10 % fetal bovine serum (BD), non-essential amino acids, glutamax (Thermo Fisher Scientific), and penicillin/streptomycin in 5 % CO2 at 37 °C. Semi-confluent cultures of Caco-2 cells were collected by treatment with Tripsisn-EDTA solution (Life Technologies) and washed with the medium. The cells were then seeded to 96-well flat-bottom microplate wells (Thermo Fisher Scientific) at a concentration of 2x104 cells/well. Flagellin solution (100 pmol/ml) was added to each well and incubated for 4 h. Culture supernatants were then collected and stored at −20 °C until use. Human interleukin-8 (IL-8) was quantified with the OptEIA kit (BD) in accordance with the manufacturer’s instructions. Assays were duplicated or triplicated and repeated three times at least. Tukey’s multiple comparison test was performed and P values less than 0.05 were considered as statistically significant.
5
Genetically Modified HeLa Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HeLa-mCAT#8 cell line that stably expresses the mouse ecotropic retroviral receptor mCAT-1 was used as the HeLa WT cell line, and its CERT1-disrupted mutant cell line was used as the HeLa CERT KO cell line [5 (link)] unless otherwise noted. To avoid unwanted overexpression of CERT cDNAs, the HeLa CERT KO/shCERT cell line was used. The HeLa CERT KO/shCERT cell line was established as described in a separate publication (Goto et al., submitted). In brief, the CERT KO cells were transfected using a retroviral vector containing a short hairpin RNA against CERT1 RNA to interfere with the expression of CERT1 mRNA. Then, a stable clone was isolated and used as the parental HeLa CERT KO/shCERT cell line. The CERS2 KO cell line (#16) was established as described previously [6 (link)]. HeLa cells were cultured in high glucose Dulbecco’s modified Eagle medium (D-MEM; 044-29765, Wako, Osaka, Japan) supplemented with 10% heat-inactivated fetal calf serum.
6
Establishment of Luciferase-Expressing Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human breast cancer MDA-MB-231/Luc cells stably expressing firefly luciferase were obtained from Cell BioLabs, Inc., (San Diego, CA, USA). Tamoxifen-resistant human breast cancer MCF-7-Luc (TamR-Luc#1) cells stably expressing firefly pGL3 luciferase were donated by Dr Kazuhiro Ikeda (Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama, Japan). Human cervical carcinoma HeLa-Luc cells stably expressing firefly pGL3 luciferase were obtained from Caliper Life Sciences Co., (Hopkinton, MA, USA).
MCF-7-Luc cells were cultured in Dulbecco's modified Eagle medium (DMEM; Wako Pure Chemical Industries, Ltd.) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 µg/ml kanamycin and 0.5 mg/ml G418 at 37°C in a 5% CO2 humidified atmosphere. MDA-MB-231-Luc cells were cultured in DMEM supplemented with 10% FBS and 100 µg/ml kanamycin at 37°C in a 5% CO2 humidified atmosphere. HeLa cells were cultured in Eagle's minimum essential medium (Wako Pure Chemical Industries, Ltd.) supplemented with 10% FBS and 100 µg/ml kanamycin at 37°C in a 5% CO2 humidified atmosphere.
MCF-7-Luc cells were cultured in Dulbecco's modified Eagle medium (DMEM; Wako Pure Chemical Industries, Ltd.) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 µg/ml kanamycin and 0.5 mg/ml G418 at 37°C in a 5% CO2 humidified atmosphere. MDA-MB-231-Luc cells were cultured in DMEM supplemented with 10% FBS and 100 µg/ml kanamycin at 37°C in a 5% CO2 humidified atmosphere. HeLa cells were cultured in Eagle's minimum essential medium (Wako Pure Chemical Industries, Ltd.) supplemented with 10% FBS and 100 µg/ml kanamycin at 37°C in a 5% CO2 humidified atmosphere.
7
Gpc3-Targeting siRNA for Cancer Therapy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse Gpc3-specific siRNA and negative control (N.C.) siRNA (Silencer Select siRNA) were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). The siRNA sequences were as follows: Gpc3 siRNA sense: 5′-GAG UCA GUC UUA GAC AUC AdTdT-3′ and Gpc3 siRNA antisense: 5′-UGA UGU CUA AGA CUG ACU CdTdT-3′. Alexa488- and Alexa647-labeled siRNAs were also synthesized by Thermo Fisher Scientific, Inc. Dacarbazine was purchased from Wako Pure Chemical Industries (Osaka, Japan). Anti-PD-1 antibody was purchased from Abcam (Cambridge, UK). Fetal calf serum (FCS) and Dulbecco’s Modified Eagle Medium (DMEM) were purchased from Wako Pure Chemical Industries (Osaka, Japan). Cell Counting Kit-8 (CCK-8) reagent was purchased from Dojindo Laboratories (Tokyo, Japan).
8
Adipose-Derived Stem Cell Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolation of ADSCs was performed according to our previous report [6 (link)]. Briefly, ~2 g to 3 g of adipose tissue was resected, washed with
Dulbecco’s phosphate-buffered saline (DPBS; Wako, Osaka, Japan), and cut into fine pieces,
which were incubated at 37.5°C for 1 hr with shaking in high-glucose Dulbecco’s modified
Eagle medium (DMEM; Wako) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich,
St. Louis, MO, USA), penicillin (100 U/ml)/streptomycin (100
µg/ml), amphotericin B (0.25
µg/ml) (100× antibiotic–antimycotic mixed stock
solution; Nacalai Tesque, Kyoto, Japan), and collagenase type I (1.0
mg/ml; Sigma-Aldrich). The digested tissue was filtered through a
sterile 100-µm nylon mesh (EASYstrainer, 100 µm; Greiner
Bio-one Japan, Tokyo, Japan), followed by centrifugation at 775 g for 5 min in 30
ml of DPBS with 1% FBS and 1 mM EDTA·3Na (Wako) (FACS buffer). The
pellet was resuspended in DMEM and seeded on culture plates with a 10-cm diameter
(Corning, Corning, NY, USA). Upon reaching 80% to 90% confluence, ADSCs were passaged on
two culture plates (10-cm diameter) using trypsin/EDTA (0.05% w/v Trypsin-0.53
mmol/l EDTA 4Na Solution with Phenol Red; Wako) after confirming the
lack of bacterial contamination. Four cell passages were performed with the same protocol,
with cultures subsequently reaching 80% to 90% confluence in a total of eight dishes.
Dulbecco’s phosphate-buffered saline (DPBS; Wako, Osaka, Japan), and cut into fine pieces,
which were incubated at 37.5°C for 1 hr with shaking in high-glucose Dulbecco’s modified
Eagle medium (DMEM; Wako) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich,
St. Louis, MO, USA), penicillin (100 U/ml)/streptomycin (100
µg/ml), amphotericin B (0.25
µg/ml) (100× antibiotic–antimycotic mixed stock
solution; Nacalai Tesque, Kyoto, Japan), and collagenase type I (1.0
mg/ml; Sigma-Aldrich). The digested tissue was filtered through a
sterile 100-µm nylon mesh (EASYstrainer, 100 µm; Greiner
Bio-one Japan, Tokyo, Japan), followed by centrifugation at 775 g for 5 min in 30
ml of DPBS with 1% FBS and 1 mM EDTA·3Na (Wako) (FACS buffer). The
pellet was resuspended in DMEM and seeded on culture plates with a 10-cm diameter
(Corning, Corning, NY, USA). Upon reaching 80% to 90% confluence, ADSCs were passaged on
two culture plates (10-cm diameter) using trypsin/EDTA (0.05% w/v Trypsin-0.53
mmol/l EDTA 4Na Solution with Phenol Red; Wako) after confirming the
lack of bacterial contamination. Four cell passages were performed with the same protocol,
with cultures subsequently reaching 80% to 90% confluence in a total of eight dishes.
9
Transfection of SOD1 variants in N2a cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used the same procedures as those used in our previous study [23 (link),36 (link)]. Briefly stated, expression plasmids (pmCherry-N1, Clontech Laboratories Inc (Mountain View, CA, USA) harboring variants of human SOD1 (wild-type (WT) or mutant (G85R)) were prepared as reported previously [23 (link),36 (link)]. The mouse neuroblastoma Neuro2a (N2a) cell line was obtained from Public Health England (London, UK). For culturing, N2a culture cells were kept in a humidified atmosphere of 5% CO2 at 37 °C and maintained in Dulbecco’s modified Eagle medium (DMEM, Wako Pure Chemical Industries Ltd., Osaka, Japan) containing 10% (v/v) fetal bovine serum (FBS, Thermo Fisher Scientific Inc., Waltham, MA, USA). N2a cells were passaged by trypsinization every 3–4 days. The transfection of mCherry, SOD1WT–mCherry, and SOD1G85R–mCherry in N2a cells was performed using Lipofectamine 2000 according to the manufacturer’s instructions (Thermo Fisher Scientific Inc., Waltham, MA, USA).
10
Isolation and Culture of Neonatal Rat Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NRCMs were isolated from neonatal Wistar rats as described previously [15 (link)]. The hearts harvested from 1–3-day-old Wistar rats were washed in PBS with 20 mM 2,3-butanedione monoxime (BDM) on ice. Then, the ventricles of the hearts were minced into small pieces and washed in wash solution (Hank’s Balanced Salt Solution with 0.08% trypsin and 20 mM BDM) for 2 h at 4 °C with stirring followed by an incubation in collagenase solution (Leibovitz’s L15 medium with 0.15% collagenase and 20 mM BDM) for 30 min at 37 °C. The suspension, tissue fragments of which were removed by filtration, was centrifuged at 100× g for 5 min at 4 °C, and the pellet was resuspended in high- glucose Dulbecco’s modified Eagle medium (DMEM; Wako, Osaka, Japan) containing 10% feral bovine serum (FBS; Gibco/Lifetechnologies, Carlsbad, CA, USA), 1% antibiotic-antimycotic mixed solution (Nacalai tesque, Kyoto, Japan) and 100 µM bromodeoxyuridine (BrdU). The cell suspension was pre-plated for 90 min twice to remove the attached non-cardiomyocytes. The non-attached cardiomyocytes were collected, seeded and cultured on culture dishes (for lucigenin assay) or coverslips coated with 1% gelatin (for DCF-DA staining or measurement of [Ca2+]i) in high-glucose DMEM containing 10% FBS, 1% antibiotic-antimycotic mixed solution and 100 µM BrdU.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!