Bacterial invasion assay with human cells - (Mar/24/2010 )
Hello,
My question has 2 parts:
1) I want to try an experiment to infect human cells (like keratinocytes) with staph or other bacteria. I don't want to incubate the cells with other cell cultures after the invasion assay (or infection), so I'm looking at incubating them in the microbiology room. There is a CO2 incubator down there, but they use it for the bacterial stuff and so is not a humidified environment. Do you think it will be okay if I incubate these cells in a dry CO2 incubator, and keep an eye on the media levels?
2) What is the purpose for growing cells in the humidified incubators? My only guess is that it prevents the media from evaporating.
Thanks!
The main reason is evaporation. And this a major issue. If water evaporates you can't trust you concentrations in the medium (osmolarity, pH ect.). The problem gets worse with time...
If you can't humidify your incubator, I would recommend that you at least fill empty wells with PBS and try to keep your cells in middle wells. Would it be possible to improvise a humidity chamber for your plate or flasks? Just rember to allow for gas-exchange.
Hope this helps
Kirsten
archercr on Mar 24 2010, 06:34 PM said:
My question has 2 parts:
1) I want to try an experiment to infect human cells (like keratinocytes) with staph or other bacteria. I don't want to incubate the cells with other cell cultures after the invasion assay (or infection), so I'm looking at incubating them in the microbiology room. There is a CO2 incubator down there, but they use it for the bacterial stuff and so is not a humidified environment. Do you think it will be okay if I incubate these cells in a dry CO2 incubator, and keep an eye on the media levels?
2) What is the purpose for growing cells in the humidified incubators? My only guess is that it prevents the media from evaporating.
Thanks!
I think it will be fine, as long as there is CO2, the cells will be happy. Yes, you can add sterile water in the other empty wells, just to prevent your medium from evaporation.