Foetal calf serum

In vitro lymphocyte proliferation in media containing rat serum compared with foetal calf serum was lower in rats fed conventional wheat compared with organic wheat in protein-energy-malnourished rats. Conventional wheat represented higher risk for lymphocyte function than organic wheat in vulnerable conditions for the rat ... 

Method. Figure 3 shows the equipment used by us for loading a culture medium with radon and decay products. Air was circulated in a closed system, driven by a membran pump (MP). The system consisted of a Ra-226 solution (Ra), a security bubble flask with water (H20), a membran bacteria filter (MF) and a second bubble flask containing 100 ml RPMI 1640 culture medium (CM). This medium contains 100 IE/ml penicilin and streptomycin and 0.75% L-glutamin. Foetal calf serum, an essential part of blood cultures, must not be added, else the airstream would develop foam. Furthermore we added a small amount of Pb(N03)2 and Bi(N03)2, about 10 ng of each, as "carriers" for the radon decay products to avoid a "wall effect". 

Antibiotics are required to prevent microbial growth consequent to accidental microbial contamination. Supplemental serum (often bovine or foetal calf serum, or synthetic serum... 

In addition to protein impurities emanating directly from the source material, other proteins may be introduced during upstream or downstream processing. For example, animal cell culture media are typically supplemented with bovine serum/foetal calf serum (2-25 per cent), or with a defined cocktail of various regulatory proteins required to maintain and stimulate growth of these cells. Downstream processing of intracellular microbial proteins often requires the addition of... 

Figure 7.1. Effect of GM-CSF on neutrophil protein biosynthesis. Human neutrophils were incubated in RPMI1640 medium supplemented with 2.5% foetal calf serum and 60 jtCi/ml [35S]-methionine, in the presence and absence of 50 U/ml GM-CSF. After 4 h incubation at 37 °C, the cells were pelleted by low-speed centrifugation. The proteins in the cell pellets were precipitated by 10% trichloracetic acid and then analysed by two-dimensional polyacrylamide gel electrophoresis, using iso-electrofocussing in the first dimension. Electrophoresis in the second dimension was performed in the presence of SDS and used a 12% acrylamide gel. Source Experiment of Becky Stringer.

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. 

The commercial exploitation of cell cultures can be gauged from the use of foetal calf serum. Even though this is increasingly being reduced as a component of cell culture media, sales increased more than 6-fold between 1984 and 1987 (Spier, 1987). The value of the market for products made from cultured animal cells has been estimated at 23 billion dollars for 1991 (Ratafia, 1987). These figures underline the importance of animal cell culture as an industrial as well as a research tool. 

Growth media, i.e. media suitable for the continued culture of established cell lines, are prepared by dilution of the 10 X stock solutions. If the 10 X stock is made with salts present (i.e. if it is obtained commercially or if it is made from powder — see below) then it must be diluted with water. If, however, it is made as described in Appendix 1, Table 13 or 14, it must be diluted with BSS as indicated in Table 15. The latter table gives methods of preparing various minimal essential media supplemented with calf or foetal calf serum and in one case also with non-essential amino acids . The various media are all based on Eagle s MEM and have been developed to improve the growth of certain cell lines or strains as indicated. 

There are a number of media available which are not based on a detailed investigation of growth requirements, but rather include crude mixtures of nutrients added to promote cell growth. These include lactalbumin hydrolysate (Appendix 1 Table 9) or yeast extract (Appendix 4) to provide an inexpensive source of amino acids or vitamins. Thus Melnick s monkey kidney media A and B (Melnick, 1955) contain lactalbumin hydrolysate and calf serum in Hanks and Earle s BSS, respectively. Chick embryo extract and tryptose phosphate broth (Appendix 1, Tables 11 and 12) are also used occasionally and their use is referred to where appropriate throughout the book. Mitsuhashi and Maramorosch mosquito cell medium contains lactalbumin hydrolysate, yeast extract and foetal calf serum in a specially developed saline (Mitsuhashi and Maramorosch, 1964 Singh, 1967). 

Add a drop of complete growth medium (e.g. Dulbecco s MEM containing 10% foetal calf serum) and cut the skin fragment into 5—10 pieces. 

Add an equal volume of complete medium. Medium F12 supplemented by up to 30% foetal calf serum has been recommended (Ham, 1965). The medium should be preincubated with 5% C02 to prevent undue alkalinity and should not be above 30 °C, otherwise the cells will attach to the walls of the vessel (Ham and Puck, 1967). 

Add 20 ml calf or foetal calf serum and 20 ml tryptose phosphate broth (Appendix 1) to 80 ml double strength medium (e.g. Glasgow MEM Appendix 1) and warm the mixture to 44°C. 

Subculture Don-C cells every 24 h in McCoy s 5a medium (Appendix 1) containing 20% foetal calf serum and 0.08% lactalbumin hydrolysate, seeding cells at 1.2 X 105/ml(3 X 104 cells/cm2). 

The gradient is formed from two solutions. Al contains either (a) 10% w/v sucrose in complete medium which has the NaCl concentration reduced by 146 mM to maintain constant osmotic pressure, or (b) 30% foetal calf serum in PBS A2 contains either (a) 2.72% w/v sucrose in compelte medium which has the NaCl concentration reduced by 40 mM or (b) 15% foetal calf serum in PBS. The volumes, and hence heights, of the two solutions must be the same (about 250 ml). 

Into B add the cell suspension (10-20 ml) in growth medium containing 10% foetal calf serum and allow this to flow into C where it will form a narrow band beneath the medium or PBS already there. 

Plate 7.5 X 105CHO cells into 5 cm dishes in Eagle s MEM lacking isoleucine and containing 5% dialysed foetal calf serum. 

After overnight incubation replace the medium with fresh medium containing isoleucine (2 X 10 6 M) and 10% undialysed foetal calf serum. 

After 8 h incubation change the medium for Eagle s MEM containing isoleucine, 10% undialysed foetal calf serum and 2 mM hydroxyurea. 

The cells grow rapidly and are hyperdiploid. They are grown in RPM1 1640 (Chapter 5 and Appendix 1) supplemented with 10% foetal calf serum and 1% glutamine. They adhere loosely to the surface but may be grown in a stationary culture or in roller bottles at 2-8 X 10 -5 cells/ml. They may be dislodged from the substratum by shaking the bottle. 

Foetal calf serum (for SV40) DEAE dextran (0.5%)... 

Sediment the cells from the supernatant and resuspend in growth medium containing 20% foetal calf serum and hydrocortisone (0.4/ig/ml) (this accelerates growth and makes colony morphology more orderly). 

Bacto-peptone 600.0 Glutathione Foetal calf serum 0.50 0-30%... 

EFC EGF EMS ESG ETC Eagle s MEM supplemented with foetal calf serum epidermal growth factor ethyl methane sulphonate Ewing sarcoma growth factor Eagle s MEM supplemented with tryptose phosphate (10%) and calf serum (10%)... 

Wash medium RPMI 1640 cell culture medium (Sigma), 10% (v/v) foetal calf serum (Invitrogen), 50 mA/HEPES buffer, pH 7.0 (Sigma), 2 mMEDTA... 

Plant products have also been detected in non-plant organisms. Morphine 1, the archetypal plant alkaloid, has in fact been shown to be a physiological plasma constituent and its production in mammals could be traced to the liver expression of the critical enzymes of its biosynthesis.17 In addition, the plant hormone abscisic acid 12 has been detected as an endogenous constituent of human brain,18 while caffeine 13 was isolated from a marine gorgonian (Paramuricea chamaelon)19 and the atisane diterpenoid serofendic acid 14, an inhibitor of the oxidant-induced mitochondrial death pathway and putative activator of mitoK(ATP) channels, has been characterised from foetal calf serum.20... 

Note For exosomes collected from urine and viruses, only proteins also identified in cell-secreted exosomes ( 1-10) are indicated. Asterisks indicate bovine proteins from foetal calf serum. 

For labeled amnion, incubate in proline/glutamine-free DMEM containing 100 Atg/ml ascorbic acid, 100 /zg/ml BAPN, 20% dialyzed foetal calf serum and 2 /rtCi/ml L-[ " C]proline incubate at 37°C on a gyratory shaker for 36 h, wash the membrane exhaustively with PBS (to remove nonincorporated radioactivity) and denude the amnion as above. 

Among the coumarins isolated from species of Artemisia (Asteraceae), esculetin (6,7-dihydroxycoumarin) and its dimethyl-ether scoparone were known to be antiproliferative on vascular smooth muscle cells. This activity was further found in some very simple mono-substituted coumarins, which were even more potent than esculetin, although less effective. In an attempt to verify its mechanism of action, esculetin was tested for interactions with PTK and PKC. The induction of membrane PTK activity by either foetal calf serum or platelet-derived growth factor (PDGF) was moderately reduced by esculetin, whereas no effect was observed against PKC [56]. 

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. 

A graphic presentation of a murine hybridoma grown in a fixed-bed bioreactor under batch mode is given in Figures 5.9.4 and 5.9.5. The medium used in these examples was Dulbecco s modified Eagle s medium (DMEM) with 4.5 g glucose, 4mM glutamine and 5% foetal calf serum (FCS). 

Complete culture medium The serum free medium is supplemented with 10% heat-inactivated foetal calf serum (Gibco, Grand Island, MY, USA). 

Final cumulative doses of 22.2-29.6 GBq (600-800 mCi), with treatment intervals of 6-9 weeks, are reported in the literature. In this study, AR42J rat pancreatic tumour cells were grown in F12-K medium (GIBCO) supplemented with 10% foetal calf serum and incubated at 37°C. Cells (2.5 x Kf cells/mL) were cultivated on a 60 mm diameter plate and exposed to 2400 or 5700 kBq/ mL of [ LuJDOTATATE for 1 h, washed twice with PBS and then cultured with cytochalasin B (Sigma, USA) at a final concentration of 2 pg/mL for 72 h at 37°C. After 72 h, cells were fixed, stained and analysed for the presence of MN in binucleated cells. 

Determination of specific binding of the labelled peptides to biological materials was carried out with D341 Med human medulloblastoma cells derived from a tumour biopsy from a patient with a cerebellar medulloblastoma, which expresses high levels of somatostatin receptors. The cells were maintained as a continuous cell line in 10% foetal calf serum and zinc option media in a humidified atmosphere (37°C, 5% CO,). 

To study the antiviral activity, Type 1 simplex herpes virus, KOS stock (HSV-Hg) and Indiana vesicular stomatitis virus (VSV) were used and HeLa cells were cultured in Eagle-modified Dulbecco medium (EMDM), supplemented w ith 10% foetal calf serum, on plates with 24 holes. Monolayers of these cells were infected with HSV-1 or VSV at 0.5 ufp/cell or 0.01 ufp/cell, respectively, and later the product to be assayed, pre-dissolved in DMSO, was added in concentrations of 10, 20, 50, 100 and 200 pg/ml. After 48 h incubation for HSV-1 and 24 h for VSV, at 37°C in CO2 atmosphere, the cytopathic CPE effect was measured on a phase-contrasting microscope [85],... 

In cultured fibroblasts, mRNA concentration increases 4-fold under the influence of serum (Wion et al., 1985 Houlgatte et al., 1986, 1989), and preliminary studies have provided some limited description of the physical characteristics of the active agent(s) (Houlgatte et al., 1989). Augmented expression of mRNA jf by cultured astrocytes and enhanced release of NGF protein into culture medium under the influence of foetal calf serum was reported by Furukawa et al. (1987) and Spranger et al. (1990). 

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