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My cells died after changing media... - (Jan/18/2010 )

Hi ,
I am really confused and sad....My cells died after changing the media...we always used FBS from Invitrogen but we changed to FBS from sigma...Do you guys think this change in FBS will affect the cells so much..?
Anyone, please help,,
Thanks so much in advance...

-Priya914-

Easily can affect the cells. Some cells are extremely sensitive to serum batches.

-Dukey-

+1

-madrius1-

Dukey on Jan 18 2010, 07:24 PM said:

Easily can affect the cells. Some cells are extremely sensitive to serum batches.

Thanks for your reply...

-Priya914-

Dukey on Jan 18 2010, 08:24 PM said:

Easily can affect the cells. Some cells are extremely sensitive to serum batches.

yup, we are always testing the batches, when we find a good one we just buy a really big stock of it.

-Radish-

Sorry to hijack, but i think it belongs to the same topic

How does one test batches of FBS?

And has anyone heard of Biowest? Apparently they claim their serum is of good quality which can give competitive prices compared to established brands like gibco.

-Psyclick-

Psyclick on Feb 13 2010, 10:44 AM said:

Sorry to hijack, but i think it belongs to the same topic

How does one test batches of FBS?

And has anyone heard of Biowest? Apparently they claim their serum is of good quality which can give competitive prices compared to established brands like gibco.



Dear Psyclick,

We buy exclusively New Zealand sourced serum because it has proved to be the best quality serum you can get for cell culture. We know this because we "Batch Test" the serum, as stated by Radish, and then purchase and store between 50-100 bottles (500ml volumes). The test we do are specific to our research and what we are after are consitant experimental results from one year to another, one decade to another. Below are the combination of test that we do in our lab and Institute. Many groups contribute to the final decision, and all experimental data is collated by me, so that there is no bias at all:-

Cell growth as defined by population doubling times
Cell Viability
The ability of J774 murine macrophages to induce iNOS enzyme
Cellular respiration rates of at least 2 of our cell lines as measured by the "Clark Electrode"
HEK 293 cells transfected with Sodium Channels.....the batches of serum will determine the amount of channels the cells express.
The concentration of intracellular Cytochrome C in intact cells (an in house method).

The results are collated by me and on every ocassion one batch is found to be superior to the others IN EVERY TEST WE DO.

We also ask specifically for batches of serum that havd LOW ENDOTOXIN LEVELS. This is specifically important to us as background endotoxin can induce some of the cellular enzymes we are interested in.


Hope this gives you a flavour of what shouls be done. It can take 3-4 weeks to get the results. However if you purchase a large enough batch then you should be doing this just once every 2 years.

p.s. remember that batches of serum have expiry dates. Do not purchase to much and allow them to expire.

Kindest regards

Uncle Rhombus.

-rhombus-