Tissue culture growth media

It has been demonstrated that the distribution of vanadium species inside the cell can depend on the form in which the vanadium is administered, as was seen in fish where a different distribution of vanadium in red blood cells (RBCs) was found depending upon whether metavanadate or decavanadate was given. In contrast to this, a similar accumulation was found in plasma and cardiac cytosol. However, the ratio of vanadium in plasma to vanadium in RBCs increased over time with metavanadate administration and remained constant for decavanadate administration. When either of the vanadium compounds was used, most of the vanadium was first found in plasma before moving into the mitochondrial fraction [9,10], Although one can know with some certainty what vanadium compound is given to an animal or put into a tissue culture growth medium, it is difficult to always know the identity of the active form inside the cell. 

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. 

Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tabacco tissue culture. Physiol Plant 15 473-497... 

Mirocha CJ, Schauerhamer B, Pathre SV (1974) Isolation, detection, and quantification of zearalenone in maize and barley. J Assoc Anal Chem 57 1104-1110 Muller R, Baier M, Kaiser WM (1991) Differential stimulation of PEP-carboxylation in guard cells and mesophyll cells by ammonium or fusicoccin. J Exp Bot 42 215-220 Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15 473-497... 

If the cells are tissne-cnlture cells intended for cell sorting, use tissue-culture medium in which the cells have been growing as the wash solution. The tissue-culture supernatant is withdrawn from the tissue-cnlture flask and filtered through a 0.22-p filter to ensure sterility. The addition of this medium helps cells recover after sorting and increases the growth of cells when they are placed back into tissue culture. 

Murashige, T. and F. Skoog, "A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures," Physiologia Plantarum 15 (1962) 473-497. 

In the process of developing an in vitro system to select phosphate starvation resistant cell lines we simultaneously selected for a line that was constitutively induced for APase excretion. Tissue cultured tomato cells were plated onto solid medium containing starvation levels of phosphate. While most cells died, we identified isolated clumps of callus capable of near-normal rates of growth. Starvation-resistant cells were used to start suspension cultures that were kept under phosphate starva-... 

CCRF-CEM T-lymphoblastoid cells (ATCC CCL 119) were cultivated in RPMI 1640 medium supplemented with L-glutamine (0.3 g/L) containing 10% bovine serum using 24-well tissue culture plates. The cells were seeded at 10s mL 1 and after a 24-h incubation period (C02 atmosphere, 37 °C) tested compounds were added at five different concentrations. The endpoint of the cell growth was 72 h following the drug addition. An appropriate aliquot from every dish was then counted (cell counter Serono 150+). The inhibitory potency of the compounds tested was expressed as IC50 values. 

When preparing a new medium the need for cell adaptation should be considered. The adjustment process for low protein media is often carried out in static cultures employing tissue culture flasks. Flowever, problems seen at the time of adaptation can be minimized in suspension cultures, since this allows subculture during the log growth phase. Cultures in suspension also allow better medium oxygenation. Flowever, under any condition and with any medium, there will always be a need for some cell adaptation to the new medium to stabilize cellular replication. As soon as a cell population is adapted for growth in a low protein medium, it is recommended that a cell bank is established. 

Anchorage-dependent cells can be grown on solid or macroporous microcarrier beads and these can then be superfused with nutrient medium in the spectrometer. The advantages of the system include direct contact between the perfusate and the cells (see below) and growth of the cells at rates comparable to those found in tissue culture dishes. A disadvantage, with solid beads, is the relatively low cell density obtainable due to the large sample volume occupied by the beads. 

To deposit cells (or tissues) at the repositories a frozen vial or a subconfluent culture in a 25 cm2 flask filled completely with growth medium is required. In the latter case the neck of the flask should be taped and the flask wrapped in pop-film in a polystyrene box or jiffy bag. The package should be kept at room temperature and delivered by the fastest route. Depositories require documentation before they will accept samples and the appropriate paperwork must be completed before the cells are despatched. 

In the appropriate culture medium, tissue explants give rise to callus tissue. Callus tissue is comprised of large, thin-walled parenchyma cells. It is similar to the undifferentiated tissue produced by plants as a repair mechanism when they are injured. In tissue culture, dedifferentiated callus can be induced to form plantlets that grow into normal plants. The induction of callus occurs when a sterile explant is brought into contact with a nutrient medium, which contains substances that initiate cell division and support growth. An explant may be a uniform piece of tissue or tissue derived from different cell types (Yeoman, 1973). Storage parenchyma tissue from Jerusalem... 

Examples of substrates released into the culture medium by plant tissue cultures are given in Table 7. The growth rate and yield can be improved by medium optimization. The products from plant tissue cultivations are either directly extracted from the cells or the medium or subjected to biotransformation and enzymatic synthesis. 

Cells are grown either in suspension in a free or immobilized form 102), or by adherence to a solid surface 100). Materials used for promoting surface-dependent cell growth are glasses, metals, plastics, carbohydrate polymers etc. the media used contain substances such as blood plasma, amniotic fluids, tissue extracts, etc.103). Recent developments in animal cell culture are aimed at the improvement of strains and culture techniques, medium optimization, and scale-up. In contrast to plant cell culture, animal cell culture has already found its technical application. Large-scale... 

Endothelial cells are maintained in a basal culture medium, such as DMEM or a proprietary medium supplied with the cells, which is supplemented with hydrocortisone (lug/mL). epidermal growth factor (EGF 100 ug/mL) bovine fibroblast growth factor (FGF 1 ng/mL), antibiotics (gentamicin and amphotericin, at 50 ug/mL) and fetal calf serum (2-10%). Alternatively, bovine brain extract (3 ug/mL) can be used instead of EGF and FGF. The cells are cultured in either standard tissue culture flasks directly on plastic or on flasks coated with collagen. The cells are grown to confluence and subcultured and reseeded in a ratio of 1 3. For all experiments primary endothelial cells should be used between passages 3 and 12. 

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