High content screening cell culture

The method comprises five individual steps (see Fig. 1), including the preparation of the transfection solutions, followed by their spotting onto a cell substrate (e.g., Lab-Tek culture dishes, Nalge Nunc International, Rochester, NY), plating of the cells onto the arrays of spotted transfection solutions, preparation of the transfected cells for functional analysis, and finally the analysis of transfected cells by high content screening microscopy. 

Fig. 1. The five steps to produce arrays of transfected mammalian cells for high content screening microscopy. 1. Preparation of the transfection solutions on an automated liquid handler. 2. Spotting of the transfection solutions with a spotting Robot, for example, ChipWriter Compact on a cell substrate, for example, Lab-Tek tissue culture dishes. 3. Plating of the cells on dishes with dried transfection solutions. 4. Preparation of samples for functional analysis, for example, immunostaining. 5. Analysis of samples by high content screening microscopy.

Arens et al. (691) reported a radioimmunoassay method for the quantitative analysis of ajmaline in plant material. Several plants were screened for their ajmaline content. The highest levels were found in R. vomitoria. Cell cultures of this plant were initiated and further optimized for ajmaline production. Tryptophan addition to the cell culture resulted only in one type of medium in an increase of alkaloid production (up to 0.04 g/liter). The RIA method also proved to be suitable in screening/ , serpentina cell clones for high production. Initial experiments showed variations in productivity over a factor 5 for cell clones obtained from one population. Cell line selection for improving alkaloid production thus seems promising. 

In an attempt to delineate the variability of serially cultured callus and cell suspension cultures derived from three cultivars of C. roseus obtained from highly uniform explants, Kurz et al. (596) screened several hundred cultures from anthers of buds at identical developmental stages. The total alkaloid content of the cell suspension cultures varied from 0 to 1.5% of dry weight. The relative amounts of alkaloids produced were found to be fairly constant for each culture and appeared to be cell line specific. 

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