Microbial cells culture media

It is necessary to estabUsh a criterion for microbial death when considering a sterilization process. With respect to the individual cell, the irreversible cessation of all vital functions such as growth, reproduction, and in the case of vimses, inabiUty to attach and infect, is a most suitable criterion. On a practical level, it is necessary to estabUsh test criteria that permit a conclusion without having to observe individual microbial cells. The failure to reproduce in a suitable medium after incubation at optimum conditions for some acceptable time period is traditionally accepted as satisfactory proof of microbial death and, consequentiy, stetihty. The appHcation of such a testing method is, for practical purposes, however, not considered possible. The cultured article caimot be retrieved for subsequent use and the size of many items totally precludes practical culturing techniques. In order to design acceptable test procedures, the kinetics and thermodynamics of the sterilization process must be understood. 

Mammalian Cells Unlike microbial cells, mammalian cells do not continue to reproduce forever. Cancerous cells have lost this natural timing that leads to death after a few dozen generations and continue to multiply indefinitely. Hybridoma cells from the fusion of two mammalian lymphoid cells, one cancerous and the other normal, are important for mammalian cell culture. They produce monoclonal antibodies for research, for affinity methods for biological separations, and for analyses used in the diagnosis and treatment of some diseases. However, the frequency of fusion is low. If the unfused cells are not killed, the myelomas 1 overgrow the hybrid cells. The myelomas can be isolated when there is a defect in their production of enzymes involved in nucleotide synthesis. Mammahan cells can produce the necessary enzymes and thus so can the fused cells. When the cells are placed in a medium in which the enzymes are necessaiy for survival, the myelomas will not survive. The unfused normal cells will die because of their limited life span. Thus, after a period of time, the hybridomas will be the only cells left ahve. 

Fructan was harvested by precipitation from the culture broth by addition of ethanol or isopropanol. Acetone and methanol can also be used. The yield and consistency of the product varied depending on the amount of alcohol added. The fructan started to precipitate at the medium/alcohol v/v ratio of 1 1.2, and the yield peaked at about 1 1.5. Further increase in the ratio hardened the fructan and made the product less fluid. Slightly less isopropanol was needed than ethanol to precipitate levan (fructan). Although most of the bacterial cells, unfermented sugars, and other solubles remained in the aqueous alcohol phase, pre-removal of microbial cells by centrifuging was needed to obtain a pure form of fructan. The product was further purified by repeated precipitation and dissolution in water, followed by dialysis or ultrafiltration. The final product was an... 

It is suggested that whereas metal ions usually penetrate poorly into a microbial cell the formation of a metal chelate may facilitate metal ion uptake from the medium in which the cell is growing, leading to toxicity. Alternatively, diffusion of the chelating agent into the cell may lead to a critical depletion of essential metal ions in the cell. Metal [Fe(II), Cu(II), Cd, Ni, Ru(II)] chelates of 1,10-phenanthroline bases are lethal to cultured mammalian cells at concentrations down to 0.1 /uM98). This cytotoxicity limits their possible medical uses to topical applications. 

For cells growing continuously in suspension, the subculture process can be performed similarly to the method used for microbial cultures. Trypsin treatment is not required and subculture is faster and less traumatic for the cells. Total medium exchange is not generally performed for these cultures since it would require a centrifugation step. Culture maintenance can be performed by dilution with fresh medium after adequate cell growth. 

The aeration method most widely used in animal cell cultures is bubble aeration, just as in microbial fermentations. This method is simple and consists of bubbling a gas stream directly into the culture medium, using a... 

Electronic noses provide new possibilities for monitor the state of a cultivation non-in-vasively in real-time. The electronic nose uses an array of chemical gas sensors that monitors the off-gas from the bioreactor. By taking advantage of the off-gas components different affinities towards the sensors in the array it is possible with the help of pattern recognition methods to extract valuable information from the culture in a way similar to the human nose. For example, with artificial neural networks, metabolite and biomass concentration can be predicted, the fermentability of a medium before starting the fermentation estimated, and the growth and production stages of the culture visualized. In this review these and other recent results with electronic noses from monitoring microbial and cell cultures in bioreactors are described. 

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