I am looking to conduct a proteomic study and need a cell line that can grow in serum free medium. Also is there a defined medium that is ideal for mass spec analysis such that the medium contents don't interfere with the MS results.
You can study with CHO and BHK 21 cell line. They can easily adapt to serum free media and easily grow.
ooo yeah, I'm overlooked. Human cancer cell line easily grown serum free media, especially human breast cancer and colon carcinoma cell line.
u may use Hela cells they jst grow like weeds ,they grow well in DMEM media or RPMI and donot innterfer with MS.
Thanks guys. Do you know of any primary lines that might grow ok in serum free conditions or chemically defined medium? I know primary cells are finicky but if anyone has experience with growing them in defined conditions, that would be useful info.
primary cell lines are preety sensitive they tend to form spheroid bodies in serum free media or low serum media, but one thing that can be done is u can supplement the media with essential growth factors like insulin , egf etc but keep in mind this growth factors are too costly , can increase your your cell culture costs is large scale.
all d best..!
1) Probably, for a proteomics study a medium can be simply replaced with a suitable serum-free one closer to the collection time (with a couple of washes to remove all traces of serum).
2) Hela cells are reported to grow very well in a serum-free media (containing all necessary nutrients and trace elements - e.g. F12 + trace elements and further MCDB series of media) supplemented with 5 additional factors - "insulin, transferrin, hydrocortisone (aldosterone), fibroblast growth factor, and epidermal growth factor":
Article Growth and Maintenance of HeLa Cells in Serum-Free Medium Su...
It appears, that hydrocortisone is the most crucial one (followed by EGF) while they will still grow (slower) without other factors: Article Replacement of serum in cell culture by hormones: A study of...
Though, the concentrations of EGF, FGF2 seem to be unphysiologically very high compared to what cells normally get from 10% serum. I'm wondering if-
- there are newer formulations with a newer (more active?) growth factor preparations;
- or whether the supplemented proteins just got adsorbed on plate walls which can be prevented by e.g. 0.1 mg/ml BSA (that's often a fatal problem when you dilute enzyme to very low concentrations and want to see the activity: Article How to prevent losses of protein by adsorption to glass and plastic
)3) A few cell lines (like CHO) can apparently grow slowly (and with some other effects) but indefinitely without any additional hormones and growth factors - presumably if they produce some proliferative minimum of growth factors themselves. A431 Human Epidermoid Carcinoma may be a good bet of a human cell line which still grows relatively well even without any protein factors: Article Epidermal growth factor inhibits growth of A431 human epider...
In that paper they have used DMEM+F12 which was likely contaminated with trace metals but today it's probably better to use MCDB 105, 201, F12+trace metals or their 1:1 combinations with DMEM.- Badges
- Science topic