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Trypsin for J774A Macrophages?


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#1 dw237606

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Posted 21 February 2011 - 01:52 PM

Can J774A mouse macrophage cells be trypsinized for subculturing or would it be better to scrape them? Currently I'm scraping but would like to use trypsin if possible. Thanks.

#2 dancedive

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Posted 24 April 2012 - 07:13 PM

Both could be used. Scraping is gonna kill a lot of cells though. trypsinization can damage the cell surface receptors (if you want to do subsequent flow analysis, this may pose a problem).

If you want to get them off the plate mildly, you can use 3 mM EDTA+10mM glucose in PBS too.

#3 rhombus

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Posted 25 April 2012 - 08:00 AM

Can J774A mouse macrophage cells be trypsinized for subculturing or would it be better to scrape them? Currently I'm scraping but would like to use trypsin if possible. Thanks.



We grow our J774 in Techne stirrer bottles because:

They grow excellently in SUSPENSION
You do not need to trypsinise the cells to set up experiments
The stirrer bottles last for 20-30 years and can be autoclaved infinetely
The cells grow to 800,000 cells/ml and are 100% viable...so large quantities can be available for big experiments

The J774 cells stick like superglue to thye TC plastic so scraping is the only alternative....but viability can go down to 40-50% i.e. you lose over half the cells..........not cost effective at all

Spend the money and buy TECHNE stirrer bottles/stirrer platforms

Hope this helps

Uncle Rhombus




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