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Any one can help me for co-culture system?


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#1 molnutri

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Posted 24 July 2014 - 12:20 PM

Hello,

 

I am trying to set up co-culture system with 3T3-L1 and C2C12.

 

I never had previous experience about co-culture.  Sure I have some experience for jsut regular cell culture.

 

Could anyone can give some general information how to set up co-culture system and what kinds of tools (dishes and where?) I should purchase.

 

Could anyone who has this two cell types coculture experience give a protocol??

 

Should I use the same media for both cells? or each their media in co culture? 

 

THANKS IN ADVANCE.

Hellen

 

 



#2 kohlmeise

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Posted 28 July 2014 - 12:25 AM

Hi! 

Sorry I don´t have experience as well, but also trying to start with co culture. Until now the idea is to culture cells in 24xmulti well dishes. Hence you have several layers one above the other. 

Furthermore I found some paper by Robert A Brown. He did some research on the field of 3D culture in collagen gels .... maybe that could be interresting for you!

 

By the way does anybody has an idea how to deal two types of cells which requires different kinds of media ? Is it possible to mix some kind of "Media Cocktail" of both required Media ?

 

Best regards !

kohlmeise

 


#3 bob1

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Posted 28 July 2014 - 12:39 AM

By the way does anybody has an idea how to deal two types of cells which requires different kinds of media ? Is it possible to mix some kind of "Media Cocktail" of both required Media ?

 

Best regards !

kohlmeise

 

The best thing to do would be to wean one onto the other medium.  This is done by slowly changing the medium into the new one over several passages.  Basically start off with (old:new) 90:10, then next passage, 80:20, 70:30... etc until your cells are totally on the new one.  It is best (mandatory) that you monitor the cells you are weaning for things like growth rate, morphology, isozyme markers etc., to ensure that they aren't changed substantially from the parent line.



#4 kohlmeise

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Posted 28 July 2014 - 12:59 AM

Ah thanks so much I will try and let you know how it works :) !

 

Best regards !



#5 SusieQ

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Posted 28 July 2014 - 10:44 PM

Maybe it's also a good idea to experiment with cell concentrations of either type, to see in what proportion the cells work best with each other? Assuming you don't already know this, of course :)






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