Minimum essential medium

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. 

The histamine- and leukotriene B4-releasing potential of the RCBPA P743 has been investigated in vitro on the RBL-2H3 mast cell line [25]. RBL-2H3 cells (with permission of Dr RP Siraganian, National Institutes of Health, Bethesda, MD, USA) were replated on Eagle s minimum essential medium and incubated for 30 minutes at 37 °C in 5% CO2 with the test-solutions. Calcium ionophore A23187 was used as a positive control. The viability of mast cells was determined by means of a quantitative colorimetric assay that is based on the abiUty of viable cells to cleave the reagent MTT [33]. 

Minimum essential medium (MEM) with glucose reduced to 0.5 mM (Gibco/In-vitrogen, Karlruhe, Germany). 

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

Figure 4. Photomicrograph of 3T3 murine fibroblasts stained with Wright s-Giemsa stain a) treated with W. floribunda agglutinin (150 ng/ml) in Eagle s Minimum Essential Medium with Hanks salts for 48 hr b) control...

The test medium used for the virucidal assays was Minimum Essential Medium (MEM) supplemented with 1-10% (v/v) heat inactivated FBS. The medium may also be supplemented with one or more of the following 10 pg/ml gentamicin, 100 units/ml penicillin, and 2.5 pg/ml amphotericin B. 

Boone et al. (1968) centrifuge cells through a discontinuous 10-20% Ficoll gradient made up in Eagle s minimum essential medium modified for suspension (i.e. lacking calcium and bicarbonate and containing 10 times the normal phosphate concentration). They use an A-1X zonal centrifuge rotor and spin for 1 h at 1000 r.p.m. at 20°C, and obtained clear separation of different cell types (HeLa and rabbit thymocytes). 

MEM Methocel minimum essential medium carboxymethyl cellulose type MC (4000 centripoi-ses) sold by Dow Chemical Co. 

Improved Minimum Essential Medium (IMEM Biofluids, Inc., Rockville, MD, USA) supplemented with 10% heat-inactivated fetal bovine serum, 100 p,g/mL streptomycin, lOOU/mL penicillin, and 2mM glutamine. 

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. 

MDCKII cells are cultured according to Cui et al. (2001). In brief, cells are cultured in minimum essential medium supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillin, and 100 pg/ml streptomycin in humidified 5 % C02/95 % air at 37 °C. For selection antibiotics are added (1 mg/ml zeocin, 800 xg/ml G418 disulfate, 500 pg/ml hygromycin B). Cells are passaged every 3 to 4 days at 1 10 to 1 15 ratios. 

The procedure of kinetic data analysis and model construction is illustrated for a hybridoma culture (cell line VO 208) in a batch system. The medium used was RPMI 1640 + 5% (v/v) foetal calf serum (PCS) + 2% (v/v) minimum essential medium (MEM) amino acids + 1% non-essential amino acids and initial glucose and glutamine concentrations of 13 mM and 4.5 mM, respectively. 

Eagle s minimum essential medium (MEM) (or equivalent alternative) with 10% foetal bovine serum (FBS), Eagle s non-essential amino acids and other supplements as necessary (see General Principles , point 4). 

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

MEM 10% Minimum essential medium (MEM, Life Technologies) supplemented with 10% inactivated fetal calf serum (FCS), 1% L-glutamine, and 0.3% sodium bicarbonate. 

Chinese hamster ovary (CHO) cells (type AA8 ATCC CRL-1859) American Type Culture Collection (Rockville, MD, USA) a-Minimum essential medium (a-MEM) from Gibco/BRL, Burlington, Canada... 

DMEM Dulbecco s modification of Eagle s minimum essential medium... 

Caco-2, C5-0, V79 cells (in Dulbecco s Modified Eagle Medium, DMEM), and Chinese hamster ovary (CHO)-Kl cells (in DMEM/F-12) at passage numbers between 30 and 50, and HepG2 cells (in Minimum Essential Medium) at passage numbers between 80 and fOO, are grown as monolayers in 80-cm culture flasks, which are harvested when they reach 80% confluence to maintain exponential growth. 

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