Dulbecco s modified Eagle

Cultures. Swiss 3T3 and A431 cells were grown in a gassed (5.5% COJ, humidified incubator in Dulbecco s Modified Eagle Media (DME) supplemented with 10% heat inactivated fetal calf serum (FCS). 

Culture of Pheochromoqtoma Cells (PC12 Cells). PC12 cells, kindly supplied by Dr. H. Hatanaka of our institute, were maintained in Dulbecco s modified Eagle s medium containing 5% heat-inactivated horse serum. 

Basic animal cell culture media, such as Dulbecco s modified Eagle s medium, generally contain ... 

EROD activity is measured in the H411E cells as follows. The cells are seeded at 7,000 cells per well in 250 xL of Dulbecco s modified Eagles culture media (Tillitt et al., 1991). After an initial incubation period of 24 hours, the cells are dosed with 5 xL volumes of eiuiched SPMD extracts (cleanup of extracts generally includes dialysis and size exclusion chromatography [SEC]) and incubated for an additional 72 hours. Sample dose is typically expressed as g-equivalents triolein or whole SPMD per mg cellular protein. Multiple exposures are performed at each of six (typically) sample concentrations, using a dilution series. Afterwards, the microtiter plates are washed three times with distilled water to lyse the cells. EROD activity (pmol mg cellular protein per min) in each sample is measured... 

Figure 4 Effect on the serum stability of the incorporation of 5% cholesterol poly(ethy-lene glycol) (PEG) into cationic lipoplexes [hpopolyamine RPR209120/DOPE1/1, ratio (mol) lipid/DNA = 10 in 150 mM NaCl]. Lipoplexes were incubated in DMEM + 10% SVF, at 37°C, aliquots were regularly sampled and monitored by dynamic diffusion. Results represent a mean between three measurements. Error bars are not presented to simplify the graph, but differences among PEG, PEG-1, and PEG-2 are significant. Abbreviations PEG, poly(ethylene glycol) DOPE, dioleylphosphatidylethanolamine DMEM, Dulbecco s Modified Eagle Medium.

For incubation, cells are preincubated for five minutes at 4°C in culture medium without buffer substances and acidified with 20 mM succinic acid, pH 5.7 [Dulbecco s modified Eagle s medium (DMEM) without sodium bicarbonate, but containing 0.6% bovine serum albumin (BSA), 20 mM 2-[N-morpholino]-ethanesulfonic acid (MES), 20 mM succinic acid, pH 5.7)]. For different methods useful for acidification of cells, see Ref. (43). 

Appropriate culture medium for the specific tumor cell line(s) under test (e.g., Dulbecco s Modified Eagle Medium (DMEM) lx liquid GlutaMAX I, 4,500 mg/L o-glucose, sodium pyruvate DMEM/F12 (1 1) lx liquid with GlutaMAX I). 

Dulbecco s Minimal Essential Medium (Dulbecco s Modified Eagle s Medium, MEM Dulbecco, DMEM)... 

Complete medium for culture of the undifferentiated ES-D3 embryonic stem cell line Dulbecco s Modified Eagle s Medium (DMEM), 20% Fetal bovine serum (FBS), 1% Nonessential Amino Acids (NEAA) (Gibco, Gaithersburg, MD), 2 mM L-Glutamine, 1% 5,000 lU/ml Penicillin/5,000 pg/ml Streptomycin, 0.1 mM P-Mercaptoethanol. 

Brandley and Schnaar [140] immobilized a synthetic nonapeptide, Tyr- Ala-Val Thr-Gly Arg-Gly-Asp-Ser, on a polyacrylamide gel, which had been prepared by a ternary copolymerization of acrylamide, bisacrylamide and the acrylic ester of. V-hydroxysuccinimide. They reported that Balb/c 3T3 mouse fibroblast cells (in Hepes-buffered Dulbecco s modified Eagle medium) adhered readily to the peptide-derivatized surfaces, even in the absence of serum, although long-term cell growth required the presence of serum. It was noticed that reference nonapeptide. Tyr-Arg-Leu-Glu-Asp-Pro-Ala-Met-Trp, which has no RGD sequence, failed to promote cell-attachment. 

Primary fibroblasts from skin biopsy samples are grown at 37°C in the presence of 5% C02 in Dulbecco s modified Eagle s medium (DMEM) supplemented with 10% foetal calf serum (FCS) and 1% penicillin/streptomycin. For passaging, confluent cells are washed with phosphate-buffered saline (PBS) and incubated for 5-10 min at 37°C in the presence of trypsin. 

Dulbecco s Modified Eagles Medium (DMEM) containing antibiotics (100 U/mL penicillin, 100 U/mL streptomycin, 0 25 pg/mL amphotericin B, 50 pg/mL genta-mycin) and 10% (v/v) fetal bovine serum (FBS). 

Reagents. The following buffers and solutions are needed for whole cell screening growth media Dulbecco s modified Eagle s medium (DMEM), 10% fetal calf serum (FCS), 100 units/ml penicillin, and 100 /ig/ml streptomycin, PBS 8 mM Na2HP04, 1.4 mM KH2P04, 137 mM NaCl, 2.7 mM KC1, pH 7.2. 

All of the cell lines are routinely maintained as monolayer in Dulbecco s modified Eagle s medium supplemented to 10% with fetal bovine serum, 100 units/ml of penicillin and 100 mg/ml of streptomycin. 

Labelling of Cells. For labelling experiments, Dulbecco s modified Eagle s medium was used, except it contained only 1% glucose. Cells were seeded at 1 to 2 x 10 > cells/75 cm2 flask in medium at 37° and allowed to attach for 20-24 hours. The medium was then replaced with fresh medium containing sodium butyrate,... 

Culture Condition. Fifteen-day-old embryos numbering 11 to 14 per litter were removed by Caesarean section (23) and placed in Dulbecco s modified Eagle s medium augmented with glucose (600 mg %), 0.23% sodium bicarbonate, 90 units/ml penicillin,... 

Dulbecco s modified Eagle s medium (DMEM) with L-glutamine and 4.5 gm/L glucose. Add penicillin-streptomycin (Gibco BRL, Gaithersburg, MD) from 100X stock. Add fetal bovine serum (FBS) to a final concentration of 10%. 

Dulbecco s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and TrypLE Express Stable Trypsin-Like Enzyme with Phenol Red (Invitrogen, Carlsbad, CA). 

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