poplietal lymphnode

Tim Axtelle taxtelle at icos.com
Wed Dec 20 12:30:02 EST 1995


In article <4b44n0$bfa at news.fhg.de>, <mmueller at ita.fhg.de> writes:

> 
> I have to prepare lymphocytes from popliteal lymph nodes. Does anyone 
> know a useful method to get enough cells from these lymph nodes. I got 
> from 10.000 up to 100.000 cells per lymph node. My problem is, I need 2.6 
> 10exp6 cells for my experiments.
> Thanks a lot
> Meike Mueller 
> 
> 

I assume that mouse is the species in question.  I am usually able to get a 
good yield using the frosted ends of frosted glass slides.  Gently rub each 
node between two frosted glass slides over a petri dish containing your medium 
of choice. Rinse the slides frequently by dipping the frosted ends in the 
medium.  When I have problems with yield, it's usually because these cells 
tend to clump.  To reduce clumping, use the frosted glass slides gently (don't 
apply a lot of pressure).  Also, once your suspension is prepared, place it in 
a culture tube and allow chunks of debris to settle for 5 minutes or so.  Then 
remove the sup. to a clean tube. You might also try suspending cells in Ca++ 
Mg++ free Phosphate buffered saline to reduce clumping.  Good luck.




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