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Any tips on how to avoid lifting cells in 96well plates


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3 replies to this topic

#1 Peniel

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Posted 20 April 2013 - 03:59 AM

Hi everyone,

I work with MTS assay for drug inhibition. I did a practise run before the real experiment and tried dosing the drug each day. Whilst changing the media I realized that some of my adherent cells tear off and form a scaffold and some get lost. A549 seems fine and does not get dislodged but some other cancer cell lines do. Has anyone experienced this and would like to give some tips? I'd appreciate it.

Thanks

#2 Tabaluga

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Posted 20 April 2013 - 07:54 AM

I suppose you pipet the fresh medium carefully and slowly at the wall of the well, or let it drop in very softly and slowly ? Cause the cells can detach through direct forces of medium being "shot" at them if one does it too quickly.

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#3 Tom M

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Posted 23 April 2013 - 01:11 PM

We have a similar problem with HEK293 cells. Using coated plates, poly-d-lysine or collagen for example, helps keep the cells attached. In addition, we don't aspirate the media but rather invert the dish and gently pat the dish on an absorbent surface to remove buffer during the washing.

Good luck!

#4 Peniel

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Posted 24 April 2013 - 03:28 AM

We have a similar problem with HEK293 cells. Using coated plates, poly-d-lysine or collagen for example, helps keep the cells attached. In addition, we don't aspirate the media but rather invert the dish and gently pat the dish on an absorbent surface to remove buffer during the washing.

Good luck!


Thanks for the responses, I will try inverting them and patting on absorbent paper. Also putting the medium in slowly.




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