Cell culture, analysis

Cell Culture Analysis Kit for the determination of phosphate and organic anions in cell cultures... 

Klebe, R. J., and Ruddle. F. H. (1969). Neuroblastoma Cell culture analysis of a differentiating stem cell system. J. Cell Biol. 43, 69A. 

Sakai, H., Okano, T., Yamada, N., Sakurai, Y. (1996). Thermoresponsive polymer surface for cell culture analysis of the surface and control of the cell attachment/detachment. In N. Ogata, S. Kim, J. Feijen, T. Okano (Eds.), Advanced biomaterials in biomedical engineering and drug delivery systems. Tokyo Springer. 

Mounting electrodes in a bioreactor is costly, and there is an additional contamination risk for sensitive cell cultures. Some other sensors of prac ticai importance are those for dissolved oxygen and for dissolved carbon dioxide. The analysis of gas exiting from a bioreactor with an infrared unit that detects carbon dioxide or a paramagnetic unit that detects oxygen (after carbon dioxide removal) has been replaced by mass spec trophotometry. Gas chromatographic procedures coupled with a mass spectrophotometer will detect 1 the volatile components. 

FIGURE 11.4 Two-way analysis of variance. Arrangement of data in rows and columns such that each row of the cell culture plate (shown at the top of the figure) defines a single dose-response curve to the agonist. Also, data is arranged by plate in that each plate defines eight dose-response curves and the total data set is comprised of 32 dose-response curves. The possible effect of location with respect to row on the plate and/or which plate (order of plate analysis) can be tested with the two-way analysis of variance. 

Auroux et al. give an up-to-date description of the application of pTAS components and systems, including cell culture and cell handling, immunoassays, DNA separation and analysis, polymerase chain reactions, and sequencing [43] (see also [44] for a description of the pTAS components and systems). 

Directed evolution relies on the analysis of large numbers of clones to enable the discovery of rare variants with unproved function. In order to analyze these large libraries, methods of screening or selection have been developed, many of which use specialized equipment or automation. These range from the use of multichannel pipettes, all the way up to robotics, depending on the level of investment [59]. Specialized robotic systems are available to perform tasks such as colony picking, cell culture, protein purification, and cell-based assays. 

For the analysis of ribosome loading in the cytosol or ER by velocity sedimentation, it is necessary to utilize a minimum of 1 to 2 X 107 cells. The techniques described here are optimized for a 75 cm2 culture flask and may be altered to accommodate other cell culture flask sizes. For time... 

The suppression and recovery of protein synthesis from DTT treatment (without cycloheximide treatment) can be monitored via metabolic pulse radiolabeling of cell cultures using [35S]-methionine and subsequent determination of radiolabeled protein content either by SDS-PAGE/ phosphor-imager analysis or liquid scintillation of tricholoroacetic acid insoluble material (Stephens et al., 2005). 

Fig. 1.2 Protein blot analysis of human therapeutic protease inhibitor (HTPI) produced in alfalfa cell cultures using different promoters and subcellular targeting peptides as shown. Equal amounts of total soluble proteins from cell cultures were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and blotted onto a polyvinyldifluoride (PVDF) membrane. Monoclonal anti-HTPI IgGs were used for detection.

---