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Cells are unhealthy

cell culture troubleshooting

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#1 Jingchun

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Posted 13 March 2014 - 11:00 AM

Hi Everybody,

 

I am culturing the fibroblast cells. My cells become unhealthy every a few generations. But my colleagues are OK. The only differences are the incubation and the FBS and Pen strep concentration. I put exactly 10% FBS and 1% Pen Strep. They just put 50 mL and 5mL respectively to the medium. For me, I put 56.1mL and 5.61mL. Why do I ask here because I am wondering the affection of a little higher FBS and Pen Strep is so much! I paid attention all of the tricky stuffs for cell culture. Somebody can give me some suggestions or something else?

 

Of course, I will try to low down the FBS and Pen Strep concentration as my colleague is doing to see what happens.

 

Thanks,



#2 bob1

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Posted 13 March 2014 - 11:52 AM

Well the logical thing to do would be to try using their conditions...

 

However, depending on the line (immortalized/transformed or not) cells do reach a point known as the Hayflick limit, where they reach will stop replicating and enter senescence, never to divide again.  This could be what you are experiencing.



#3 CellApplicationsInc

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Posted 17 March 2014 - 09:41 AM

Since your coalleague's cells cultured at the same time are ok, I don't think the problem is that your cells are reaching senescence. I also don't think those extra microliters of FBS and P/S would make a big difference. I think the devil is in those  "tricky stuffs". Is your colleague more experienced with cells? Can you ask him/her to watch over your shoulder when you are working with the cells and tell you exactly what (s)he would do different?







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