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.