The cell culture medium was performed. The preparation of 1 l of MEM supplemented with 10% FBS required 9.5 g MEM (GE Healthcare, USA), 2.2 g NaHCO3 (Merck, Germany), 100 mL fetal bovine serum (Gibco, New Zealand) and 900 mL deionized water. The MEM was gently mixed in 1000 mL beaker and steriled filtered through 0.22 μm filter upper cups (Jet Biofil, China) into 1000 mL duran bottle (Duran, Germany). The culture medium was kept at 4 °C, and warmed at 37 °C before use. The preparation of 1 l of DMEM supplemented with 10% FBS required 13.4 g DMEM (GE Healthcare, USA), 3.7 g NaHCO3 (Merck, Germany), 100 mL fetal bovine serum (Gibco, New Zealand) and 900 mL deionized water. The DMEM was gently mixed in 1000 mL beaker and steriled filtered through 0.22 μm filter upper cups (Jet Biofil, China) into 1000 mL duran bottle (Duran, Germany). The culture medium was kept at 4 °C, and warmed at 37 °C before use.
Dulbecco modified eagle medium (dmem)
Manufactured by GE Healthcare
Sourced in United States, Austria, Germany, United Kingdom, Australia, Switzerland, Japan, France, China
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture media formulation developed by Dulbecco and Freeman. It is a widely used basal medium that provides essential nutrients and growth factors to support the in vitro cultivation of a variety of cell types. The core function of DMEM is to maintain cell viability, enable cell growth, and support cellular metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (60 mentions)
Fetal bovine serum (fbs), by GE Healthcare (57 mentions)
Penicillin, by Thermo Fisher Scientific (25 mentions)
Penicillin streptomycin, by GE Healthcare (24 mentions)
Streptomycin, by GE Healthcare (24 mentions)
Penicillin, by GE Healthcare (23 mentions)
Streptomycin, by Thermo Fisher Scientific (21 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (20 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (17 mentions)
L glutamine, by GE Healthcare (17 mentions)
Fetal bovine serum (fbs), by Merck Group (15 mentions)
Hyclone, by GE Healthcare (12 mentions)
L glutamine, by Thermo Fisher Scientific (9 mentions)
Rpmi 1640 medium, by GE Healthcare (8 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (7 mentions)
Ficoll paque, by GE Healthcare (7 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (7 mentions)
Mcf 7, by American Type Culture Collection (7 mentions)
Rpmi 1640, by GE Healthcare (7 mentions)
Roswell park memorial institute (rpmi), by GE Healthcare (6 mentions)
338 protocols using dulbecco modified eagle medium (dmem)
1
Preparation of Cell Culture Media
2
Integrin-Mediated Cell Adhesion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HT1080 cells derived from a human fibrosarcoma were obtained from the European Collection of Animal Cell Cultures, Porton Down, UK. These were maintained in a humidified incubator with 5% CO2 at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM, Sigma–Aldrich) containing 10% fetal bovine serum (Sigma–Aldrich) and 1% streptavidin/penicillin (Life Technologies). Prior to cell adhesion experiments, HT1080s were detached from the cell culture flasks with 0.05% trypsin/0.02% EDTA (GE Healthcare), washed and re-suspended in serum free DMEM.
Collagen films were BSA blocked as for platelet adhesion analysis then 100 μl of HT1080 cells were added at a density of 5 × 105 cells/ml in serum free DMEM containing either 5 mM MgCl2 or 5 mM EDTA. After incubation at 37 °C/5%CO2 for 40 min loosely bound cells were removed with 3 × 200 μl PBS washes. Bound cells were detected using the phosphatase substrate as for platelet adhesion. Integrin mediated HT1080 attachment was derived by subtracting attachment in the presence of EDTA from attachment in the presence of Mg2+. Values indicate means of quadruplicate measurements ± standard deviation.
Collagen films were BSA blocked as for platelet adhesion analysis then 100 μl of HT1080 cells were added at a density of 5 × 105 cells/ml in serum free DMEM containing either 5 mM MgCl2 or 5 mM EDTA. After incubation at 37 °C/5%CO2 for 40 min loosely bound cells were removed with 3 × 200 μl PBS washes. Bound cells were detected using the phosphatase substrate as for platelet adhesion. Integrin mediated HT1080 attachment was derived by subtracting attachment in the presence of EDTA from attachment in the presence of Mg2+. Values indicate means of quadruplicate measurements ± standard deviation.
3
Matrigel Invasion Assay with BaP and U0126
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Matrigel invasion assay was performed using a 24-well Transwell chamber (Corning Incorporated, Corning, NY, USA). A total of 5×104 cells in DMEM (GE Healthcare Bio-Sciences) without FBS were seeded into the upper chamber with Matrigel. A total of 4 µM BaP or 20 µM U0126 were added to the upper chamber. A total of 600 µl DMEM supplemented with 10% FBS (both GE Healthcare Bio-Sciences) were added to the lower chamber. After 24 h of incubation at 37°C, non-migrated cells on the upper side of the membrane were removed with cotton swabs. Cells on the lower surface of the membrane were fixed with pure methanol at room temperature for 10 min and stained with crystal violet at room temperature for 30 min. The number of invaded cells was counted in six randomly selected fields. Relative ability of invasion was calculated by normalizing with control.
4
Amino Acid Starvation and Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For amino acid starvation, cells were rinsed and incubated with amino acid-free DMEM (25 mM glucose; FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan) supplemented with 10% dialyzed FBS (GE Healthcare, Tokyo, Japan) for 1 h. After the treatment, cells were rinsed and stimulated with amino acid-containing DMEM supplemented with 10% FBS for 10 min and 1 h. The formulation of amino acid-containing DMEM was as follows (in g/L): L-Arginine, 0.084; L-Cystine, 0.0626; L-Glutamine, 0.584; Glycine, 0.03; L-Histidine, 0.042; L-Isoleucine, 0.105; L-Leucine, 0.105; L-Lysine, 0.146; L-Methionine, 0.03; L-Phenylalanine, 0.066; L-Serine, 0.042; L-Threonine, 0.095; L-Tryptophan, 0.016; L-Tyrosine, 0.10379; L-Valine, 0.094.
5
Cell Culture Techniques for Diverse Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat liver hepatoma MrArdle-RH7777 and human osteosarcoma U2OS were obtained from the American Type Culture Collection. Human lung fibroblasts IMR-90 were obtained from Coriell Cell Repositories at the National Institute on Aging, Coriell Institute for Medical Research, Camden, NJ. IMR-90 and U2OS were maintained in DMEM plus 10% fetal bovine serum (FBS; GE Healthcare, Chicago, IL). MrArdle-RH7777 were maintained in DMEM plus 20% FBS (GE Healthcare). Cells were kept in a 37°C incubator with 5% CO2. Transfection of DNA constructs into IMR-90 U2OS cells was performed using Lipofectamine 2000 as detailed in the manual provided by Invitrogen. Oleic acid treatment was used at 0.75 mM oleic acid complexed to fatty-acid–free bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO). Ascorbate treatment was used at 0.25 mM ascorbic acid (Sigma) and 1 mM ascorbic-2-phosphate (Sigma-Aldrich). This concentration was in addition to the amount of ascorbic acid present in FBS (0.08 mM), as supplied by the manufacturer. Doxycycline (Sigma-Aldrich) was used at 1 µg/ml. BFA (Sigma-Aldrich) was used at 10 µg/ml.
6
Vero Cell Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Toxicity towards Vero cells was determined as described previously [44] (link) with modifications. Briefly, strains were grown to exponential phase in TSB (Difco), then diluted 1∶100 in 5 ml TSB, and incubated for 20 h at 37°C with agitation (180 rpm). Next, 100 ul of 8-fold to 512-fold DMEM (GE Healthcare) diluted cell free culture supernatants of the TSB-grown strains were added to washed confluent Vero cell monolayers in 100 ul DMEM/10% FCS in 96 well plates in triplicates. For each experiment fresh culture supernatants were produced and equal growth of the bacterial cultures was confirmed by OD600 readings. After 48 h of incubation at 37°C, supernatants were analyzed for LDH release by means of the CytoTox96 Non-Radioactive Cytotoxicity Assay (Promega) according to the manufacturer’s protocol. All values shown are corrected by the background reading of the diluted culture supernatants. In all assays, 256-fold diluted E. coli culture supernatants were determined as best discriminative and are therefore shown.
7
Cytotoxicity Assay for Bacterial Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Toxicity towards Vero cells was determined as described previously [17 (link)]. Briefly, strains were grown to exponential phase in TSB (Difco), then diluted 1:100 in 5ml TSB and incubated for 20h at 37°C with agitation (180rpm). Next, 100ul of 8-fold to 512-fold DMEM (GE Healthcare) diluted cell free culture supernatants of the TSB-grown strains were added to washed confluent Vero cell monolayers in 100ul DMEM/10% FCS in 96 well plates in triplicates. For each experiment fresh culture supernatants were produced and equal growth of the bacterial cultures was confirmed by OD600 readings. After 48h of incubation at 37°C, supernatants were analyzed for LDH release by means of the CytoTox96 Non-Radioactive Cytotoxicity Assay (Promega) according to the manufacturer’s protocol.
8
Viability Assay of AZD-1775 on Breast Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BT549, HS578T, and MDA-MB-231 cell lines (ATCC) were maintained in RPMI (ThermoFisher Scientific, Waltham, MA, USA), DMEM, and DMEM (GE Healthcare Life Sciences, Hyclone, Logan, UT, USA) media respectively. All media was supplemented with L-glutamine and 10% FBS (ThermoFisher Scientific, Gibco, Waltham, MA, USA). For viability assay, cells were seeded at 5000 cells/well in 96-well plates. After 24 h, the media was removed and replaced with media containing AZD-1775 at various concentrations between 0 and 3.2 µM, DMSO was used as vehicle and given in control wells. Growth was monitored every 4 h to ensure control wells reached but did not exceed 95% confluence. After approximately 72 h of treatment for each cell line, Cell Titer Glo® (Promega, Madison, WI, USA) viability assay was performed as suggested by manufacturer. Luminescence values were obtained from VICTOR Multilabel plate reader (PerkinElmer, Waltham, MA, USA) and normalized to control well before plotting. Graphing and IC50 determinations were done using Prism 8 software (GraphPad, San Diego, CA, USA).
9
Cell Culture Protocols for MDCK, Phoenix, and HEK293
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDCK cells, kindly provided by Dr. N. Simmons (Newcastle University) [25] were cultured at 37°C and 5% CO2 in DMEM (catalog no. E15-810, GE Healthcare, Glattbrugg, Switzerland) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich, Buchs, Switzerland), 2 mM L-glutamine and 1% non essential amino acids (catalog no. M11-003, GE Healthcare).
Phoenix amphotropic retrovirus producer cells, kindly provided by Dr. G. Nolan (Stanford University) [26] , were cultured at 37°C and 5% CO2 in DMEM (catalog no. E15-843, GE Healthcare) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 2 mM L-glutamine and 1% non essential amino acids.
Human embryonic kidney cells (HEK293), kindly provided by Dr. A. Odermatt (University of Basel) [27] were grown in DMEM (catalog no. E15-810, GE Healthcare) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 2 mM L-Glutamine and 1% non essential amino acids at standard cell culture conditions (37°C, 95% relative humidity and 5% CO2).
Phoenix amphotropic retrovirus producer cells, kindly provided by Dr. G. Nolan (Stanford University) [26] , were cultured at 37°C and 5% CO2 in DMEM (catalog no. E15-843, GE Healthcare) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 2 mM L-glutamine and 1% non essential amino acids.
Human embryonic kidney cells (HEK293), kindly provided by Dr. A. Odermatt (University of Basel) [27] were grown in DMEM (catalog no. E15-810, GE Healthcare) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 2 mM L-Glutamine and 1% non essential amino acids at standard cell culture conditions (37°C, 95% relative humidity and 5% CO2).
10
Evaluating Mg-10Gd Degradation with Liposomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An in vitro immersion test calculating the weight loss after 72 h immersion under cell culture conditions (37 °C, 5% CO2, 20% O2, humidified atmosphere) was performed as described in Nidadavolu [53 (link)] in order to determine the impact of liposomes on the degradation of the Mg-10Gd material. Material discs 1.5 mm in height and 1 cm in diameter were cleaned utilizing 20 min sonication in n-hexane (Merck, Darmstadt, Germany), 20 min sonication in acetone (Merck, Darmstadt, Germany), and 3 min sonication in 100% ethanol. Lastly, specimens were sterilized and dried in 70% ethanol under sterile conditions [54 (link),55 (link)].
The discs were placed in Dulbecco’s modified eagle’s medium (DMEM, Life Technologies, Darmstadt, Germany) + 10% FBS (fetal bovine serum, PAA Laboratories, Linz, Austria): αMSH-SM-liposome (25:1 v:v), and in DMEM and DMEM 10% FCS: PBS (phosphate-buffered saline) (25:1 v:v). The Mg concentration released into the immersion medium was measured utilizing atomic absorption spectrometry (Agilent 240 AA, Agilent, CA, USA), as described [54 (link)]. The immersion assay was also performed with pure Mg using the same methodology.
The discs were placed in Dulbecco’s modified eagle’s medium (DMEM, Life Technologies, Darmstadt, Germany) + 10% FBS (fetal bovine serum, PAA Laboratories, Linz, Austria): αMSH-SM-liposome (25:1 v:v), and in DMEM and DMEM 10% FCS: PBS (phosphate-buffered saline) (25:1 v:v). The Mg concentration released into the immersion medium was measured utilizing atomic absorption spectrometry (Agilent 240 AA, Agilent, CA, USA), as described [54 (link)]. The immersion assay was also performed with pure Mg using the same methodology.
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!