Cell/tissue culture animal

The masterpiece of modem biotechnology is the generation of valuable flavouring substances by means of cell tissue cultures derived from plant and animal organisms. In chapter 2.2 the most important processes for the preparation of and the production with cell cultures are described. 

The implementation of mathematics into biology, physiology, pharmacology, and medicine is not new, but its use has grown in the last three decades as computer speeds have increased and scientists have begun to see the power of modeling to answer scientific questions. A conference was held in 1989 at the National Institutes of Health called Modeling in Biomedical Research An Assessment of Current and Potential Approaches. One conclusion from that conference was that biomedical research will be most effectively advanced by the continued application of a combination of models—mathematical, computer, physical, cell, tissue culture and animal—in a complementary and interactive manner. ... 

Farm animals produce recombinant proteins less expensively than bacteria or cells in culture because the farm animals produce large volumes of milk containing up to 5 g/L of recombinant protein. In addition, modifications to the proteins that can be performed only by mammalian cells are made by the cells of the mammary gland. Therefore, numerous pharmaceuticals that previously could only be made by cells in culture or extracted from human tissue or blood are being produced by lactating farm animals. 

Animal cell cultures that are initiated from cells removed directly from the animal are called primary cultures (Figure 2). Primary cultures include both explant cultures (i.e., cultures initiated from small pieces of intact tissue), as well as cultures initiated from preparations of individual or dispersed cells (obtained from intact tissue by mechanical or proteolytic dismption). Nerve fiber explant cultures in blood plasma were among the earliest types of tissue cultures (Harrison, 1907). Cells grow out from such tissue explants and form a single layer of cells completely filling the tissue culture vessel surface. Such cell cultures are called confluent monolayers. Confluent monolayers can then be treated with trypsin, so as to remove the individual cells from the culture vessel surface. The resulting cell suspension is then transferred into other culture containers, so that more viable monolayer... 

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. 

Many vimses, both DNA and RNA containing, will cause cancer in animals. This so-called oncogenic achvity of a vims can be demonstrated by the observahon of tumour formahon in inoculated experimental animals and by the ability of the vims to transform normal tissue culture cells into cells with malignant characteristics. These transformed cells are easily recognizable as they exhibit such properties as rapid growth and frequent mitosis, or loss of normal cell contact inhibition, so that they pile up on top of each other instead of remaining in a well-organized layer. 

A more general example fiom virus vaccine produetion is the rigorous examination of tissue cultures to exclude contamination with infectious agents fiom the sonrce animal or, in the cases of human diploid cells or cells fiom eontinuous cell lines, to detect... 

The term cell culture refers to the growth of isolated cells in vitro, whereas tissue culture is a term used to describe the growth of not only isolated cells, but also isolated tissues or organs. Both these terms are often used to describe the growth of animal and human cells in culture. Advances in medical research have largely driven animal cell culture development. 

As well as overcoming many of the inherent problems associated with agriculture, plant tissue culture also offers a number of advantages over conventional animal cell culture methods currently being applied to produce biopharmaceutical proteins commercially [8], As plant culture media are relatively simple in composition and do not contain proteins, the cost of the process raw materials is reduced and protein recovery from the medium is easier and cheaper compared with animal cell culture. In addition, as most plant pathogens are unable to infect humans, the risk of pathogenic infections being transferred from the cell culture via the product is also substantially reduced. 

The natural fluorescence of CTC and its derivatives has been used extensively to determine small amounts of CTC in biological materials. Kohn (86) showed that the fluorescent complex formed by CTC with calcium ions and barbital could be extracted from animal tissues into an organic solvent and then measured spectrofluorometrically. The intense fluorescence of anhydro-CTC was used by Hayes and DuBuy (87) to determine CTC in animal tissues, tissue culture cells, and bacteria. Poiger and Schlatter (88) extracted CTC from biological material into ethyl acetate as the CTC-calcium trichloroacetate ion pair. The fluorescence of the antibiotic was then enhanced by the addition of magnesium ions and a base. 

The bone becomes depleted of calcium salts when the urine is acidic over a relatively long period. This was shown by Goto (17) who fed rabbits large doses of hydrochloric acid. He then showed that urinary calcium loss occurred in concert with a marked reduction in mass of the skeletal system, and also that the total non-fat dry weight of bone decreased,implying a loss of bone matrix. A dose-dependent, dietary acid induced loss of labelled calcium from rat bone has been reported by Thorn and his coworkers (18). They demonstrated that in response to graded doses of ascorbic acid, cells in tissue culture, and bones in whole animals fed such doses were depleted of the labelled calcium. 

Studies of the enzyme content of cells frequently involve the use of coarse tissue samples of either animal or plant origin. In such cases some preliminary dissection of the tissue may be necessary to isolate the relevant tissue components and remove unwanted structural material such as collagen, cellulose, etc., before moving on to the more critical disruption of the cells. Sometimes it is possible to use the technique of tissue culture to provide pure cell preparations for subsequent studies. 

Tissue culture, more frequently used as cell culture, enables animal and plant cells to be cultured in large numbers by techniques comparable to those used in microbiology but, because of the fragile nature of the cells, does require special cultural conditions. The culture media used must supply all the essential factors for growth, such as a wide range of amino acids, nucleotides, enzyme co-factors as well as indeterminate factors that can only be supplied in special products, e.g. foetal bovine serum. The environmental conditions must be carefully controlled, particularly pH, and this is frequently maintained by culturing in a bicarbonate buffer system and a carbon dioxide saturated atmosphere. 

Only after all these exhaustive tests are a few candidates selected for pre-clinical in vivo studies using animal disease models. The current approach is to perform as many tests as possible based on tissue cultures or cell-based assays, as they are less costly and provide results more readily. At the end of this long process is the availability of selected drug candidates with sufficient efficacy and safety required for human chnical trials. 

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