F-12 medium

For further work, serum has been replaced by insulin and transferrin, and a routine assay for retinoids in serum-free medium is performed as follows (Breitman et al., 1980a) HL-60 cells are seeded at a density of 2 x 10 cells per ml in defined medium, which is a 1 1 mixture of Dulbecco s modified Eagle s minimum essential medium and Ham s F-12 medium supplemented with 3 x 10 M selenium dioxide, insulin (5 xg per ml), and transferrin (5 ig per ml). Retinoids are added to the HL-60 cells at the start of the assay, using ethanol as the vehicle final concentration of ethanol does not exceed 0.1%. The cells are incubated for 4 or 5 days, and differentiation is then measured by NBT reduction (Collins et al., 1979) of cytospin slide preparations. Results are expressed as percentage of NBT positive cells. Approximately 4-8% of the cells will differentiate spontaneously in the absence of added retinoid. As with the F9 system, dose-response curves have been determined for a large number of retinoids, and some of the more significant results are shown in Table III. 

Cells are maintained in appropriate culture media (e.g., for CHO cells, Hams F-12 medium is often used, Gibco-BRL, Rockville, MD, cat. no. 11059-029, supplemented with 10% FCS), with appropriate antibiotic and selection agents in the case of engineered ceil lines. Endogenous receptors or transiently transfected cell lines, of course, do not generally need selection. Stocks are described in detail in the case of CHO cells. 

HEK 293 cells are grown in Dulbecco s MEM / Nut Mix F-12 supplemented with 2 mM L-glutamine, 100 U/mL penicillin-streptomycin and 10% heat-inactivated fetal bovine serum (FBS complete medium). 

Hum F-12,[ 2] DM-160 and DM-170, etcJ l The MIT groupf 1 created the High-GEM (High Growth Enhancement Medium) in which fhictose replaces glucose as the energy source to achieve a 3- to 4-fold decrease in the accumulation of lactic acid. These basal media are now commercially available. 

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. 

Adherent cells were maintained in DMEM/F-12 culture medium supplemented with 10% heat-inactivated fetal bovine serum and 100 U/ml penicillin-100 /.tg/ml streptomycin at 37 °C and under 5% CO2. In our experiments, VNOf90 cells derived from rat vomeronasal organs were used. Two days before use, cells which were in a confluent state with a density of about 10 /cm were released from wells using 0.25%i trypsin-1 mM EDTA. A drop of cell solution (about 10 of the cells) was plated on the center of 35 mm tissue culture dishes and incubated for about an hour. After 1 h incubation, cells were weakly attached on dishes and the dish was then filled with culture medium. The cells were only located at the center of the dish when we performed mRNA extraction with the AFM. This procedure is quite important to decrease the probability of mRNA contamination, because dead or floating cells are one of the causes of contamination. Two hours before use, the cells were rinsed with culture medium without FBS two or three times, and the dish was filled with FBS-free culture medium. 

In this section, the application of the sensing mechanism described in Section 3.1 to an in vitro environmental model containing interfering cell culture medium is described. In this experiment the cell culture medium was used in place of deionized water, that is, 0.5 ml DMEM F-12 culmre media containing 1% fetal bovine serum (FBS) was mixed with 5.5 ml AuNPs stock solution to obtain a final AuNPs concentration of 2.25 X 10 NPs ml with pH 6.85. Additionally, to adjust the ion concentration, as was done previously, 100 pi of 10 mM FeCl3 at pH 2.90 was added into the mixture. Finally, 1 ml of 15 mM GSH at pH 3.44 was added to mediate the AuNPs. All optical characterizations were performed within 6 h of solution preparations. 

Thermal black Carbopack C Carbopack F Sterling HT Sterling FT 6-12 Medium boiling compounds, structural and spatial isomers... 

Linoleic acid and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma Chemical Co. (St. Louis, MO). Dulbeccos Modified Eagle s Medium/Hams F-12 nutrient mixture (DMEM/F12) containing 2 pM linoleic acid, fetal bovine serum (FBS), trypsin-EDTA, penicillin-streptomycin, selenium and transferrin were purchased from Gibco BRL (Rockville, MD). Other chemicals used were of reagent grade. 

Figure 14.12 Medium peeling with dermasanding. (A) and (B) Preoperative (C) and (D) postoperative 1 week (E) and (F) postoperative - after 3 months...

Fig. 12.5. Jominy test on a steel of medium hordenobility. M = martensite, B = boinite, F = primary ferrite,...

Restrictive ring, carbon ring 20 High temp. (700°F), medium pressure 12-51, 12-53... 

Aqueous dioxan (50%) has been used as a medium for bromination with acidified hypobromous acid and de la Mare and Harvey195 showed that, with perchloric acid as catalyst, the bromination of toluene followed the usual kinetic equation (89). At 25 °C, in ca. 0.0013 M hypobromous acid, the average value of fc2/[H+] for toluene (0.008-0.15 M) was 21.7 and for benzene (0.0011-0.016 M) was 0.60, so that ktoWtnc/kbtaztae was 36.2. The bromination of f-butylbenzene196 and biphenyl197 gave k2/[H+] = 7.25 and 7.52, and hence relative rates of 12.1,... 

Xie, S., Svec, F., and Frechet, J.M.J., Rigid porous polyacrylamide-based monolithic columns containing butyl methacrylate as a separation medium for the rapid hydrophobic interaction chromatography of proteins,. Chromatogr. A, 775, 65, 1997. 

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