I'm currently searching for vendors to provide L-cystine dihydrochloride for use in plant and animal cell culture media. I'm attempting to obtain a version with no more than 0.5% loss on drying; however, the specifications listed by vendors vary widely. The vendor I'd like to order from has a maximum of 5.0%. I've had a hard time finding much information on whether the loss on drying value will have any significant impact on the performance of the various media we work with. The concentration of L-cystine dihydrochloride tends to vary between 13 mg/L and 150 mg/L, although there are a couple rarer formulations that have less than 1 mg/L or over 1000 mg/L. For reference, the animal cells in my lab include BHK, CHO, and VERO attachment cells; and Sp2/0 and Sf9 suspension cells. We also perform micropropagation of various plants (Hosta spp., Vitis spp., Solanum tuberosum, Tradescantia zebrina, etc). Any insight would be greatly appreciated.
There is a work (in Russian) which shows the analysis the dynamics of amino acids in the process of culturing cells that produce recombinant glycoprotein. All the studied amino acids were divided into several groups according to their dynamics. The first group included amino acids whose concentration decreased (serine, threonine, valine, methionine, proline, isoleucine, leucine, tryptophan, phenylalanine, cystine, lysine). It was assumed that these amino acids are necessary for cell growth and prolongation of their viability. An increase in the concentration of the most rapidly consumed amino acids (cystine and serine) in the initial nutrient medium allowed to increase the yield of biomass. The second group consisted of amino acids whose concentration remained virtually unchanged (hydroxyproline, histidine, and tyrosine). They were probably in excess in the original nutrient medium. Lowering their concentration had virtually no effect on the yield of biomass and target protein. The third group included amino acids accumulated during cultivation (glycine, glutamic acid, alanine). Increase in concentration in the medium alanine inhibited the consumption of proline, which slowed the growth of CHO cells. It is shown that a decrease in the concentration of alanine and an increase in the concentration of proline in the initial nutrient medium allowed to increase the yield of biomass in relation to the initial one.
So the effect of amino acids loss on the cell culture media should not be sighnificant.
please, anyone has idea about the optimal parameter for cultivation of BHK cells in Bioreactor ( Agitation, DO, CO2 and PH).
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