Eagles medium

Mammalian cells culture media Eagle MEM 4 Atmospheric air... 

Primary Neuronal Rat Cell Culture-. The cortical rat neuronal cells were harvested by performing cervical disarticulation on 1-day-old rat pups. The cells were placed in 12- and 24-multiwell plates with a 1 1 coated surface of poly-L-lysine (Sigma Aldrich) and deionoized water (DI) to enhance cell affinity for attachment. The neuronal cells were cultured in Neuronal Culture Media (NCM) comprised of Fetal Bovine Serum (FBS), Horse Seram (HS), glucose solution, glutamine, antibiotics. Ham s F12-K (ATCC) and Basal Media Eagle s as previously described (Daniel and DeCoster, 2004). 

Basal Medium Eagle According to Hanks (BME Hanks)... 

Table 5.5 Composition of Eagle s Minimum Essential Medium (Eagle, 1959)...

Metastasis 2. Medium for tumor cell culture, e.g., Minimal Essential Medium Eagle (MEM) (Gibco, Invitrogen) 3. 10X Trypsin/EDTA dilute to final concentration lx with PBS (Sigma-Aldrich) 4. Standardized tumor cell suspension, radiolabeled 5. Hank s balanced salt solution, Ca++ and Mg++ free (CMF-HBSS) (Sigma-Aldrich, H-2387) 6. Trypan blue stain 7. Mouse vice 8. Heat lamps... 

Inject 2.5 ml growth medium (Eagle s MEM plus 10% calf serum and heparin at 10 U/ml), together with some air into the peritoneum. Do not puncture the gut. The abdomen balloons up sealing the site of injection. 

Prepare a basal layer of 7 ml of medium (Eagle s Glasgow modification containing tryptose and calf serum) containing 0.5% Difco Bacto agar in a 6 cm dish. 

ATCC BHK cells BME BPL BRL BSA BSS BUdr CHO cells CMF CMP, CDP, CTP American Type Culture Collection baby hamster kidney cells basal medium (Eagle) /J-propiolactone Buffalo rat liver bovine serum albumin balanced or buffered salt solution bromodeoxyuridine Chinese hamster ovary cells calcium- and magnesium-free BSS cytidine monophosphate, diphosphate, triphosphate... 

PSN solution (penicillin 10,000 units/mL, streptomycin 10 mg/mL, and nystatin 1250 units/mL) or Minimum Essential Medium Eagle with non-essential amino acids (MEM-NAA) medium supplemented with L-glutamine (2 ni /). 5% FCS, and 0.05% PSN solution, or other as required for each cell type. 

MEM Minimum Essential Medium Eagle with Earle s salts, without L-glutamine. 

Basal medium (Eagle) containing 25mM HEPES (Life Tedmologies, Paisley, Scotland). 

As far as is known, resistance in trypanosomes always takes the same form the parasite so modifies the chemistry of its surface that the drug is no longer taken up but remains behind in the medium (Yorke et aL 1931). On the other hand, a susceptible trypanosome can accumulate in its interior a 500 times greater concentration of arsenic than exists in the external medium (Eagle, 1945). 

In the procedure for the surface test (313), the vims is grown in a monolayer of baby hamster kidney cells and incubated in Eagles medium supplemented with tryptose phosphate broth and calf semm. After separation of the vims from the cells by sonification and centrifugation, amounts of the suspension containing 3 x 10 plaque-forrning units are dried on coversHps. The inoculated coversHps are placed in 5 ml of the disinfectant for 1, 5, or 10 min, then rinsed, sonicated, and assayed. 

Historically, the development of animal cell culture systems has been dependent upon the development of new types of tissue culture media. Mouse L cells and HeLa cells were developed using a balanced salt solution supplemented with blood plasma, an embryonic tissue extract, and/or serum. In 1955 Eagle developed a nutritionally defined medium, containing all of the essential amino acids, vitamins, cofactors, carbohydrates, salts, and small amounts of dialyzed serum (Table 1). He demonstrated that this minimal essential medium (MEM) supported the long-term growth of mouse L and HeLa ceils. Eagle s MEM was so well defined that the omission of a single essential nutrient eventually resulted in the death of these animal cells in culture. 

Eagle s MEM with serum rapidly became a standard growth medium for culturing animal cells in vitro. A number of variations of this medium were developed, including Dulbecco and Vogt s modified Eagle s essential medium (DMEM) (Table 2). DMEM contains nonessential as well as essential amino acids. The essential amino acids and vitamins are at concentrations which are significantly elevated as compared to MEM. 

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. 

HeLa S3 (ATCC CCL-2.2) cells, a clonal derivative of the parent HeLa line (ATCC CCL-2), which are adapted to grow in suspension and therefore more suitable for large biomass production, are used for the preparation of human cell extract. The cells are maintained in suspension culture in Coming 850 cm2 Polystyrene Roller Bottles at 37° at a concentration of 3 to 6 x 105 cells/ml in Eagle s Minimum Essential Medium Joklik Modification (Sigma) supplemented with 10% Fetal Bovine Serum (Invitrogen) in the presence of 5% C02. 

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

Seek T75 plastic tissue culture flasks with a minimum of 2.5 x 106 cells in 120ml of Eagle s medium containing 20mM L-glutamine 0.88g l-1 sodium bicarbonate 20 mM HEPES 50 pg ml-1 streptomycin sulphate 50IUml 1 benzyl-penicillin and 7.5% fetal bovine serum. The flasks are incubated for 18-24 h at 37°C in a C02 incubator to establish monolayer cultures. 

Prepare treatment medium containing various concentrations of test compound 19.7 ml of Eagle s medium (without serum) plus 300 pi of stock concentration of compound in a preferred solvent (e.g., water, ethanol, DMSO, etc.). The final concentration of solvent other than water should not exceed 1% v/v. Normally a range of 0-5000 pg ml-1 (final concentration) is covered. For a sparingly soluble compound, the highest concentration will be the lowest at which visible precipitation occurs. Similarly, if a compound has a marked effect on osmolality, concentrations should not be used that exceed 500 milliosmoles (mosm) per kg. In addition, a pH range of 6.5-7.5 should be maintained. 

The trypsinized cultures are counted and a sample is assessed for survival as for the cytotoxicity assay. In addition, an appropriate number of cells are reseeded for estimation of mutation frequency at the day 8 expression time. The cells are transferred to roller bottles (usually 490 cm2) for this stage. The bottles are gassed with pure CO2, the tops are tightened and the bottles are incubated at 37°C on a roller machine (approximate speed 0.5-1.0 rev min-1). Usually 106 7 8 9 viable cells are reseeded in 50 ml of Eagle s medium containing serum, but more cells are required at the toxic dose levels. 

However, fullerene C60-modified surface is adequate for the adhesion and normal growth of cells in culture. Cells of the line MA-104 in the Eagle-MEM medium formed on fullerene film a normal monolayer. Cellular viability was assessed with the resazurin (Alamar Blue) reduction test. The dye resazurin is reduced by mitochondrial dehydrogenases of viable cells into the fluorescent product resorufin (maximum X = 530 nm, max X = 590 nm). The intensity of... 

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