Serum vs cell adhesion

9 posts / 0 new
Last post
angga2cool
angga2cool's picture
Serum vs cell adhesion

Dear all kind scientists,

Could you all please help me.. In what concentration of FBS, one cells still can adhere? Im planning to treat my cells in serum free condition.. however im worry that my cells would not adhere to the plate before i treat. So i would like to use FBS as minimum as possible..

Thankx for your suggestion..

Guy Sovak
Guy Sovak's picture
Hi,

Hi,
Depending what type are your cells. HeLa needs 10%.
Some other i.e. BHK require 5%.
Guy

Omai
Omai's picture
You might want to pretreat

You might want to pretreat the dish with poly-D-lysine as an adherence factor. This may help your cells stick in serum free media. I pretreat a well for 15min. in 0.01% poly-L-lysine at room temp., remove the lysine and let the dish dry, then add 293T cells for transfection.

Omai

g a
g a's picture
It's a tricky question ......

It's a tricky question ....... although many of us are unaware of the answer but the active component oof serum that helps in cellular attachment is  feutin. most of the cells require 10% fbs which contains enough feutin  to support the cellular attachment.
There are reports showing culture of granulosa cells in as low as 1%fbs too.
But I dont think any of us ever calculated how much feutin would be enough??????

samm
samm's picture
ARGERINE wrote:

ARGERINE wrote:

It's a tricky question ....... although many of us are unaware of the answer but the active component oof serum that helps in cellular attachment is  feutin. most of the cells require 10% fbs which contains enough feutin  to support the cellular attachment.
There are reports showing culture of granulosa cells in as low as 1%fbs too.
But I dont think any of us ever calculated how much feutin would be enough??????

Do you have a reference for that?
Normal levels of fetuin in adult human serum is pretty low, with the exception of some cancers.

samm
samm's picture
Also, I've had MDCK cells

Also, I've had MDCK cells attach (with some coaxing) at 0.25% FBS, but they required 1% + supplements to actually do anything at all. Most primary immune cells (e.g. T cells and macrophages) are perfectly happy in 5% FBS.

heehawmcduff
heehawmcduff's picture
I would tend to just do a

I would tend to just do a serial dilution (perhaps from 10-0%) of decreasing serum concentrations in a culture plate and see at what point the cells stop sticking down.
Regards

Jason King
Jason King's picture
 

 
It is quite common to condition your cells over time to grow in lower concentrations of serum  for particular purposes. The success of this will depend on the cell type you are working with. There are also alternatives to serum which can be used for example in the production of monoclonal antibodies from hybridoma cell lines. As already mentioned there are various coatings such as collagen, fibronectin, vitronectin and matrigel which could help support adhesion of your cells in low serum conditions - if they were not able to adapt naturally.
 
Maybe your institution has a copy of this in the library:
 
Analytical Biochemistry Volume 102, Issue 2, 1 March 1980, Pages 255-270 

Methods for growth of cultured cells in serum-free medium
David Barnes and Gordon Sato

References and further reading may be available for this article. To view references and further reading you must purchase this article.
 

heehawmcduff
heehawmcduff's picture
And as mentioned before in

And as mentioned before in the forum you could test to see if your cells would grow in a completely serum free environment but with the supplementation of a serum-substitute such as Ultroser G
http://www.pall.com/biopharm_48468.asp