Minimal essential medium

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. 

MEL Metabolic equivalent level MEM Minimal essential medium MG Myasthenia gravis MGSA Melanoma-growth-stimulatory activity MHC Major histocompatibility complex... 

Fig. 2.2 Stability of IgCi monoclonal antibody added to sterile plant and animal cell culture media. ( ) Murashige and Skoog (MS) medium (A) Dulbecco s minimal essential medium (DMEM) with 10% serum and (A) serum-free Ex-cell 302 medium. The error bars indicate standard errors from triplicate flasks. (Reproduced with permission, from B. M. -Y. Tsoi and P. M. Doran, Biotechnol. Appi. Bio-chem. 2002, 35, 171-180. Portland Press on behalf of the IUBMB.)...

A confluent monolayer of Madin-Darby canine kidney (MDCK) cells was grown in 96-well plates. Serial tenfold dilutions in minimal essential medium were prepared from the aliquots of allantoic fluid taken from the irradiated specimen. These dilutions were applied to MDCK cells and incubated for 48 h at 36 °C in 5% C02. The cells were then washed two times for 5 min with phosphate buffered saline (PBS) and incubated for 1 h with 100 pi of 0.5 mg/ml solution of 3-(4,5-dimethyl-thiazolyl-2) 2,5-diphenyltetrazolium bromide (MTT, ICN Biochemicals Inc., Aurora, Ohio). After lh, the colored deposit was dissolved in 100 pi DMSO, and optical density in the wells was measured on plate reader Victor 1420 (Perkin Elmer, Finland). Based on the data obtained, the infectious titer of the vims was determined as a decimal logarithm of reciprocal to the dilution of the specimen causing destruction of 50% of cells. The inhibiting action of irradiation was evaluated by decreasing the vims titer. 

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

Minimal essential medium (MEM) with Earl s salts and L-glutamine (Sigma M4655). Note complete MEM is used during fibroblast culturing, but the in vitro probe medium (IVPM see below) is used during the 72-h incubation described in the procedure. 

Preparation of coverslips for scanning electronmicroscopy. Coverslips were pretreated as described by Campbell and Williams (32). These coverslips were placed in a petri dish (35 mm) onto which the cell suspension was plated out. Dulbeco s minimal essential medium plus 20% fetal calf serum was added to the dish. Each dish was then incubated at 37° in 10% CO2. Each coverslip was recovered, washed in Hanks B.S.S. (Gibco) and then immersed in Hanks B.S.S. containing 4% glutaraldehyde for at least 4 hrs. 

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... 

Fig. 4.2. Growth cycle of mouse L929 cells. Mouse L929 cells were inoculated into 2 oz medical flat bottles (5 X 10s cells in 5 ml Eagle s Minimal Essential Medium containing 10% calf serum). The medium was changed every two days. Bottles were incubated for 60 min with 2 /xCi (6-3H)-thymidine (80 /iCi/pmol) at a final concentration of 5 gM after which they were harvested by trypsinisation, their...

The mouse L-929 fibroblast line was cultivated in Eagle s Minimal Essential Medium (MEM) plus 10% calf serum. Cells were seeded in 100-mm-diameter cell culture plates at 4 x 106 cells per plate and allowed to become established for 24 hr prior to use. After the monolayer was washed, 10 mL of an agar overlay consisting of 2% Bacto-Agar and 2 x MEM was added to each plate and allowed to solidify. 

Fig. 1. The 500-MHz Hahn spin-echo H NMR spectra of a cell culture medium, Dulbecco s minimal essential medium, before (A) and after (B) reaction with 400 iiM cisplatin for 24 hr at 37°C, and after (C) reaction with Pt(en)Cl2l. Note the disappearance of the singlet at 2,14 ppm for the S-methyl of L-Met, which has formed Pt-Met complexes, giving new peaks at —2.7 ppm. (Adapted from Ref. 23.)...

Vero cells are maintained in minimal essential medium (MEM)... 

Cells are maintained and propagated under standard conditions (5% CO2 in Eagle s minimal essential medium containing 5% PCS, 2 mM glutamic acid and 50 xg gentamycin). Two days prior to the experiments, the cells are seeded into 12 or 24 well Costar (Cambridge, MA) plates at a density of 10 and 5 x 10 cells/well, respectively in the same medium at 37 °C in a humidified atmosphere with 5%C02. 

A sterile, well-buffered medium, such as Dulbecco s minimal essential medium (DMEM) without phenol red or sodium bicarbonate, supplemented with 20 mM HEPES and set at pH = 7.4 at 37°C, (which is 7.58 at room temperature because of temperature coefficient of HEPES). Store sterile at 4°C. 

Delbecco s minimal essential medium, complete serum free medium and fetal bovine serum (Cellgro, Kansas City, MO). 

Cell Culture. BSC-1 cells were grown in minimal essential medium (MEM) with Earle salts supplemented with 10% fetal bovine serum (FBS) and 1.1 g/1 sodium bicarbonate. Cells were passaged according to conventional procedures by using 0.05% trypsin plus 0.02 wt% ethylenedi-aminetetraacetic acid (EDTA) in a HEPES-buffered balanced salt solution. Tissue culture flasks were incubated at 37°C in a humidified 3% CO2 - 97% air atmosphere. Total cell counts were made using a Coulter counter equipped with a 100-pm orifice and microscopic cell count. 

RPMI 1640 media, Ham s F12K media, and Eagle s Minimal Essential Medium, MEME, respectively. All cell lines were grown in a tissue culture incubator supplied with 5% C02 and maintained at 37°C. 

In Vitro Cell Culture. Mouse leukemia L1210 cells (designated K25) were grown in RPMI 1630 medium supplemented with 20% heat-inactivated fetal calf serum. V79 Chinese hamster lung cells were grown in o-MEM medium with 5% fetal calf serum In 7.5% CO2. The human cell lines were grown in Eagle s minimal essential medium with 10% fetal calf serum. 

Tissue-culture media endotoxin-free Dulbecco s modified Eagles medium (DMEM), minimal essential medium-alpha (MEMa) or Hepes-buffered RPMI-1640 supplemented with 2 mM r-glutamine, and heat-inactivated fetal bovine serum (FBS, to 10% final concentration) (see Note 1). Store at 4 °C. 

Minimal essential medium (MEM) with Earle s salts without L-glutamine (Invit-rogen, Carlsbad, CA). 

Hanks Solution. For tissue culture (sterile, phenol red added as pH (indicator) (Sigma H8389). This replaces the more expensive MEM (minimal essential medium). 

Recommended formulations employ commercially available components for Eagle s Minimal Essential Medium (MEM). Suggested supplements collectively enhance retention of diploidy, facilitate in vitro adaptation of malignant drivatives, minimize serum toxicity, and stabilize ionic and osmological relationships between cell and medium. These modifications are recommended when single-serum supplemented MEM Eagle s and... 

Medium 80 ml MEM-S (Eagle s Minimal Essential Medium modified for suspension culture), 20 ml 20% fetal calf serum, 1 ml L-glutamine, 100 U. penicillin, and 100 fjLg streptomycin per milliliter total. 

Cell A mutant K1 cell line of Chinese hamster ovary cells (CHO cells) which lacked the enzyme dihydrofolate reductase (DHFR) was used as a host cell (3). It could not grow without supplements, such as thymidine, glycine, and purine. The expression of DHFR was used as a selective maker of genetically modified cells. A culture medium was composed of a-Minimal Essential Medium (Sigma, St. Louis, MO) supplemented with 50 U/L Penicillin and 50U/L Streptomycin (Whittaker Bioproducts, Inc. Md.) and 5% dialyzed fetal bovine serum (Whittaker Bioproducts, Inc. Md.)... 

There have recently been reports (Lowenberg et al, 19941 Callen et al, 1995) indicating that passivation of Ti-6A1-4V in HNO3 increases the release of all three constituent elements in a culture medium (a-Minimal Essential Medium with 15% foetal bovine serum and 10% antibiotics). Table 9.17 exemplifies the results obtained for titanium ions, for three periods of immersion of three days each. The level of Ti is significantly reduced throughout the 9-day experimental period. 

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