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Establishing human neural stem cells


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#1 purple_sashe

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Posted 24 February 2014 - 03:01 AM

Hi

 

A week ago I defrosted and seeded a new vial of human neural stem cells. I have made up the media with appropriate growth supplements and followed the suppliers protocols, changing the media daily. The stem cells are taking longer than anticipated to become 90% confluent. The suppliers protocol says that they should be confluent within 4 - 7 days. Today is day 7 and they are still about 40% confluent. I have no infection in the flask and as I say I have followed the suppliers protocol exactly. Ideas anyone?

 

Thanks



#2 Tabaluga

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Posted 24 February 2014 - 03:16 AM

So it is an adherent culture, are your flasks coated with something ?

Which medium do you use ?

Are the cells checked for mycoplasma ?


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 purple_sashe

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Posted 24 February 2014 - 03:45 AM

Yes its an adherent cell culture, flask coated with poly l ornithine and laminin as per protocol. I haven't tested for micoplasma, but the cells are direct from manufacturer so one would hope that they weren't infected! smile.png



#4 Tabaluga

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Posted 24 February 2014 - 04:52 AM

This may be a stupid question, but when you say you followed the recommendations exactly, does this include the CO2 percentage ? I used to work with glioma stem cells (suspension, though) and the CO2 percentage can make quite a difference in growth.

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.

 





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