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Serum free medium [View Printable]
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gsovak
Group: Member
Posts: 544
Joined: Jan 25, 2005
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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 |
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| Posted Mar 21, 2006, 18:12 PM |
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macbride
Group: Member
Posts: 24
Joined: May 02, 2005
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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.
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| Posted Mar 22, 2006, 0:43 AM |
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parvoman
Group: Member
Posts: 253
Joined: Jul 28, 2005
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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. |
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| Posted Mar 22, 2006, 16:51 PM |
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gsovak
Group: Member
Posts: 544
Joined: Jan 25, 2005
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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. |
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| Posted Mar 22, 2006, 14:29 PM |
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R Bishop
Group: Admin
Posts: 300
Joined: Jan 17, 2006
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You will be fine. I routinely starve cells overnight (16hrs) in serum-free media. 5 hrs should be no problem.
RB |
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"To those who would tear the world down: We will defeat you. To those who seek peace and security: We support you. And to all those who have wondered if America's beacon still burns as bright: Tonight, we proved once more that the true strength of our nation comes not from the might of our arms or the scale of our wealth, but from the enduring power of our ideals: democracy, liberty, opportunity and unyielding hope."
-- Barack Obama, Nov. 4, 2008
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| Posted Mar 23, 2006, 16:22 PM |
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frasermoss
Group: Admin
Posts: 704
Joined: Feb 22, 2005
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"Opportunity is missed by most people because it is dressed in overalls and looks like work". Edison
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| Posted Mar 24, 2006, 15:39 PM |
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samm
Group: Moderators
Posts: 409
Joined: Mar 03, 2005
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| 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 |
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| Posted Mar 25, 2006, 0:29 AM |
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gsovak
Group: Member
Posts: 544
Joined: Jan 25, 2005
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Thank you all for the support and help
Guy |
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| Posted Mar 27, 2006, 17:12 PM |
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