THP-1 serum free cells - (Apr/08/2008 )
As part of a larger project, I am planning to undertake some cell culture research re Zinc transporters. The cell-line I would like to work with is human THP-1, but I am having difficulty locating THP-1 cells that have been grown in media free from fetal calf serum (FCS) [I am sure most of you are aware of the ethical and technical reasons for eliminating FCS, particularly now that there are so many serum-free options available]. Does anyone have any THP-1 cells that have been adapted to serum free media, or have any suggestions as to who I could contact to source some?
Many thanks 
Meika
Many thanks
Meika
Dear Meika,
Cell culture is an artificial system and should only be used as a starting point and not a definitive model. Serum free has come some way in the past 30 years. However the basis of most products used in serum free media's is the same......hydrocortisone, transferrin, Selenium etc etc.
These additives are extremely dirty compounds and as such have massive effects on the cells themselves.
Can I ask you a question.......how are you going to grow your cells in relation to a CO2 incubator and atmospheric conditioning?
I await your reply.
Regards
Rhombus
Dear Rhombus
Thank you very much for taking the time to reply. My intended cell culture work is a small initial part of a larger human supplementation project, so it wouldn't be used as a definitive model but simply to give some indication of useful transcipts to measure when working with fresh human blood. Since posting my question I have been in touch with researchers who have successfully adapted THP-1 to serum free media, but unfortunately these groups no longer have supplies of these cells (nor do the major cell repositories stock them). To use THP-1 cells, I would therefore need to use FCS as part of the adaptation process (albeit in decreasing amounts). For ethical reasons, I have decided to rework my protocol and use PBMCs instead so the issue appears to have resolved itself.
To answer your question, I was intending to grow the cells using similar conditions to those in relevant published papers (which from memory commonly seems to involve incubating at 37°C and 5% CO2 in a humidified atmosphere - we have labs set up to do this). Why do you ask? Did you have some advice regarding appropriate conditions?
Kind regards
Meika
Thank you very much for taking the time to reply. My intended cell culture work is a small initial part of a larger human supplementation project, so it wouldn't be used as a definitive model but simply to give some indication of useful transcipts to measure when working with fresh human blood. Since posting my question I have been in touch with researchers who have successfully adapted THP-1 to serum free media, but unfortunately these groups no longer have supplies of these cells (nor do the major cell repositories stock them). To use THP-1 cells, I would therefore need to use FCS as part of the adaptation process (albeit in decreasing amounts). For ethical reasons, I have decided to rework my protocol and use PBMCs instead so the issue appears to have resolved itself.
To answer your question, I was intending to grow the cells using similar conditions to those in relevant published papers (which from memory commonly seems to involve incubating at 37°C and 5% CO2 in a humidified atmosphere - we have labs set up to do this). Why do you ask? Did you have some advice regarding appropriate conditions?
Kind regards
Meika
Dear Meika,
Most published papers containing cell culture state:-
"The cells were cultured at 37oC in a humidified 5 % CO2 Incubator"
Serum free conditions is only one variable you are addressing in this model. Atmospheric oxygen is 21%......so all these cells are grown in hyperoxic conditions.....what effect does this have on the cells?
In our own lab we have a COY hypoxic chamber where cells can be grown and passaged at levels of oxygen approaching tissue oxygenation levels over long periods of time (many months). There can be massive differences between experimental results obtained by just varying oxygen levels.
Regards
Rhombus