I have used with or without in different labs, and I am forming an opinion currently, but I am interested in other opinions too.
Dear Robert,
Streptomycin, in combination with penicillin, is used in a standard antibiotic cocktail to prevent bacterial infection in cell cultures. We used it routinely for the mass growth of procyclic Trypanosoma brucei. Penicillin inhibits the formation of peptidoglycan cross-links in the bacterial cell wall of gram negative bacteria, while streptomycin is a protein synthesis inhibitor. It binds to the small 16S rRNA of the 30S subunit of the bacterial ribosome, interfering with the binding of formyl-methionyl-tRNA to the 30S subunit. As such the antibiotic cocktail prevents the overgrowth of your cell culture by bacteria. Trypanosomes have a very flexible metabolism and are capable to switch on and off their mitochondrial metabolism, especially in the presence of glucose-rich media used for the cultivation of these parasites. It may be possible that streptomycin has an inhibitory effect on mitochondrial protein synthesis of trypanosomes as well. While studying mitochondrial metabolism of Trypanosoma brucei, I repeatedly experienced that certain mitochondrial functions such as cyanide-sensitive respiration, were repressed after growth in the presence of streptomycin. When I left out streptomycin, this property returned. Thus for those studying mitochondrial aspects of trypanosomes be careful with the use of this antibiotic.
Fred
Dear Robert,
Streptomycin, in combination with penicillin, is used in a standard antibiotic cocktail to prevent bacterial infection in cell cultures. We used it routinely for the mass growth of procyclic Trypanosoma brucei. Penicillin inhibits the formation of peptidoglycan cross-links in the bacterial cell wall of gram negative bacteria, while streptomycin is a protein synthesis inhibitor. It binds to the small 16S rRNA of the 30S subunit of the bacterial ribosome, interfering with the binding of formyl-methionyl-tRNA to the 30S subunit. As such the antibiotic cocktail prevents the overgrowth of your cell culture by bacteria. Trypanosomes have a very flexible metabolism and are capable to switch on and off their mitochondrial metabolism, especially in the presence of glucose-rich media used for the cultivation of these parasites. It may be possible that streptomycin has an inhibitory effect on mitochondrial protein synthesis of trypanosomes as well. While studying mitochondrial metabolism of Trypanosoma brucei, I repeatedly experienced that certain mitochondrial functions such as cyanide-sensitive respiration, were repressed after growth in the presence of streptomycin. When I left out streptomycin, this property returned. Thus for those studying mitochondrial aspects of trypanosomes be careful with the use of this antibiotic.
Fred
Hi, we use it routinely for both procyclic and bloodstream T. brucei culture. I had cultures with or without and saw no growth difference, but I was not doing any highly sensitive analysis like metabolomics analysis for eg.
Hi Fred, thanks for the advice. I was considering the effects that these antibiotics could have on the cells, and it is certainly useful to know that streptomycin has an effect on translation in the trypanosome mitochondrion, not surprising given its endosymbiotic origin. At Lancaster, the Ginger and McKean labs did not use antibiotics, and we managed fine without them. I am considering whether to stop using them again for my current experiments. Best wishes, Rob
Hi Catarina, thanks for your reply. It's interesting to note that you do use antibiotics as well, I was not sure if this was just more popular in US labs, than in the UK. Cell culture procedures also seem to recommend not to use antibiotics except in primary cell cultures, although I suspect most labs use them for mammalian cultures routinely too.
- Badges
- Science method