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FBS store

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FBS store

Postby jasmina » Sat Oct 31, 2009 5:11 am

hello everyone! i made a mistake yesterday, after i thawed FBS at 37℃,actually i should inactivate it at 56℃ for 30mins, but our water bath could not heat-up quickly, so at last i forgot it and left it at room tempreture overnight. i am worry about it now, can i use it for cell culture?
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Postby JackBean » Sat Oct 31, 2009 5:25 am

Fetal bovine serum?

So, you wish to inactivate it and now you worry, it won't be working? I don't get this...
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Postby jasmina » Sat Oct 31, 2009 5:36 am

To JackBean:
thank you for your reply. yes, it's fetal bovine serum, i mean after inactivation, can i use it for cell culture or not? i am worry about it now.
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Postby JackBean » Sat Oct 31, 2009 6:08 am

I would guess so. In my opinion, it's there only as nutrient source, right? In that case, inactivation should not matter (especially at 56°C)
http://www.biolib.cz/en/main/

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Postby jasmina » Sat Oct 31, 2009 6:55 am

To JackBean
thank you very much! yes, it's used as nutrient source only, at first, i think pehaps some components changed after long time storage at room tempreture. (my senior query me seriously today, although i don't think it matters, but i have to confirm it)
thousands thanks!
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Postby JackBean » Sat Oct 31, 2009 7:10 am

In my opinion, the most relevant nutritients are the proteins, which will serve as nutrition no matter their state. The low-molecular components, if any, will be only minor.
(Are you adding some supplements? Or what do you grow on it actually?)
http://www.biolib.cz/en/main/

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Postby jasmina » Sat Oct 31, 2009 7:44 am

To JackBean
I did not add anything to it, i used it for BMM culture.
HyClone Characterized Fetal Bovine Serum

here are the information about hyclone FBS, and we used SH30396.03.

Thermo Scientific HyClone Characterized Fetal Bovine Serum is characterized FBS and is comparable to the highest quality product from other suppliers.




Product Detail



Ensure superior cell culture performance with this cost-effective, high quality FBS from HyClone. It is lower in price than Defined FBS, but meets the requirements of most discriminating cell culturists.
--------------------------------------------------------------------------------

Filtered through three sequential 100nm (0.1µm) pore-size rated filters
U.S. and Canadian sourced

--------------------------------------------------------------------------------
Additional assays performed (subject to change without notice): Gamma Globulin, Alkaline Phosphatase, Lactate Dehydrogenasse, Glutamic Pyruvic Transaminase (SGPT), Glutamic Oxaloacetic Transaminase (SGOT), pH, Total Protein, Albumin, Blood Urea Nitrogen, Creatinine, Total Bilirubin, Sodium, Potassium, Calcium, Chloride, Inorganic Phosphorus, Osmolality, Iron, Total Iron Binding Capacity (TIBC), Percent Saturation, Glucose, and IgG.
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Re:

Postby JackBean » Sat Oct 31, 2009 8:21 am

jasmina wrote:To JackBean
I did not add anything to it, i used it for BMM culture.
HyClone Characterized Fetal Bovine Serum

bone marrow derived macrophages? (just write it in full name, I'm not familiar with these abbreviations ;) )
I meant, did you add something t the media? I guess you didn't culture it on pure FBS, did you?
http://www.biolib.cz/en/main/

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Postby jasmina » Sat Oct 31, 2009 8:52 am

To jackBean
I am sorry i misunderstood your meaning, yes , i added hyclone FBS to hyclone alfa-media ( added antibiotic too) ,and used it for Bone marrow derived macrophage .
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Postby biohazard » Sat Oct 31, 2009 11:09 am

Inactivation of FBS is done to prevent unwanted complement activity while still retaining the growth-supporting functions of serum. Serum is not primarily a nutrient source in most cell culture media, it's main function is to facilitate cell growth by providing required cytokines and other growth factors the cells need.

Nutrients, such as amino acids and glucose, are added to the medium separately. For many cell culture systems heat inactivation is not required, but if it is, you can do it afterward even if the FBS has been at RT overnight. If the FBS is sterile, it should be ok to use after inactivation.

Leaving a sterile FBS at room temperature for longer time (especially in direct light) may cause some molecules to degrade over time, so the growth-supporting function of the serum gets poorer. However, since the cells are kept in an incubator at +37C or so anyway, one day at +23C is unlikely to cause any detectable effects on the serum's function.

Edit: Typofix 041109
Last edited by biohazard on Wed Nov 04, 2009 7:25 am, edited 2 times in total.
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Postby jasmina » Wed Nov 04, 2009 2:33 am

To biohazard
thank you very much for your explanation. I have done osteoclast formation with this FBS these days, results show that it's OK FOR cell culture.^_^
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Re: FBS store

Postby MJD » Thu Nov 05, 2009 9:12 am

First of all, like other have said, I think the serum should be fine left out overnight.
However, I'll state that I have cultured cells for a long time without having "inactivated" the FBS serum. Additionally, I believe the term inactivation is a misnomer and the proposed function is to release growth factors that are typically bound and sequestered by binding protein. A key growth factor in FBS is IGF-1, which is bound to IGF-1 binding protein. I still would question the necessity of heating the serum. In labs, often times certain techniques are learned and continue to be used out of habit. The only way such a method is extinguished is if there is highly convincing reason that significantly impacts the lab.

A side note, some cells are very hardy and can grow in the absence of serum for days at a time.
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