Hi all,
I am working with BHK cells. For one of my experiments I need to use a DMEM/F12 without serum (usualy I use 5%FBS). Any one have got an expireance with this kind of studies?
How long can I grow the cells without serum.
Guy

I've grown cells in low serum media (1-2%) for a couple of days. I think I remember reading that serum starvation can be used to synchronize cells, so I probably wouldn't go longer than 2-3 days. I'm not sure what sort of experiment you're doing, but you could also try Opti-MEM, which is media optimized for low serum use.

It depends on why you need to be serum free and for how long.

When infecting cells in vitro with adenoviruses we usually remove the serum for the first hour or so to make infection more efficient (or to enable us to use less virus).

It is possible to slowly get your cells used to low serum levels. This is good if the cells need to be low or no serum for a few days (maybe because the cells are expressing and secreting a recombinant protein into the medium).

Otherwise there are serum substitutes (which are generally quite pricey) which get used for growing up hybridoma cells - again because the Abs are purified from the culture medium.

I need to do an asay in which I use anti Insulin receptor Ab. The serum contains insulin so I dont want it inside the Medium for at least 5Hours.
Guy

parvoman said:

It depends on why you need to be serum free and for how long. When infecting cells in vitro with adenoviruses we usually remove the serum for the first hour or so to make infection more efficient (or to enable us to use less virus). It is possible to slowly get your cells used to low serum levels. This is good if the cells need to be low or no serum for a few days (maybe because the cells are expressing and secreting a recombinant protein into the medium). Otherwise there are serum substitutes (which are generally quite pricey) which get used for growing up hybridoma cells - again because the Abs are purified from the culture medium.

You will be fine. I routinely starve cells overnight (16hrs) in serum-free media. 5 hrs should be no problem.

RB

I dont know what your apllication fully is, but have you looked into these medias and resources?

http://www.invitrogen.com/content.cfm?pageid=3817

http://www.invitrogen.com/content/sfs/brochures/SpecialtySFMediaforCC.pdf

http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Product_Lines/Serum_Free_Media.html

http://www.cellgro.com/shop/customer/home.php?cat=314

http://www.medibix.com/CompanySearch.jsp?cs_choice=c&clt_choice=t&treepath=16945&stype=i

gsovak said:

Hi all, I am working with BHK cells. For one of my experiments I need to use a DMEM/F12 without serum (usualy I use 5%FBS). Any one have got an expireance with this kind of studies? How long can I grow the cells without serum. Guy

Hi! I've done HEK293Ts in serum free conditions (following washing off of 10% serum media) for 16 h with no problems - though cell cycle pogression comes to a near total halt by then.
Also, I have routinely transferred B cell hybridomas to serum-free media - but that has to be done in a phased manner (1:2 or 1:3 dilutions, until the serum is all gone) - and in "pure" SFM, the cells grow for 6-8 days before they deteriorate.
-sam

Thank you all for the support and help
Guy