```NK cells viability post sorting on FACSAria

cdoxsee@mmm.com cdoxsee at mmm.com
Tue Jul 20 16:15:40 EST 2004






We routinely sort NKs.	I suggest sterile PBS.	I was told never to use
FACSFlow on the
Aria.  Our viability usually looks pretty good (>80%) right after sorting.
We sort at 70 psi
with a 70 micron nozzle.  What buffer the cells are in during sorting and
what solution they
go into can also affect the cells' happiness/viability.  We put the cells
in HBSS with 2%
FBS & 25 mM Hepes.  And sort into FBS.	After 48 hours we still have pretty
good
viability (around 60%) in culture.

Hope this helps,

Christie Doxsee
Immunologist
Drug Discovery, Pharmacology
Pharmaceutical Division
3M Center, Bldg. 270-2S-06
St. Paul, MN 55144

651-733-4906
fax: 651-737-5886
cdoxsee at mmm.com


|---------+------------------------------->
|	  |	      Frederic.Grosjean at is|
|	  |	      rec.unil.ch	  |
|	  |				  |
|	  |	      07/20/2004 12:11 PM |
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  |	 Subject:  ```NK cells viability post sorting on FACSAria			 
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Dear Flowers,

We tried to sort NK cells on our FACSAria last week and unfortunately the
viability of
the sorted population rapidely dropped down to zero two days after sorting.
Did anyone
sort NK cells on a FACSAria ? What conditions were used ? Could the 70 psi
be too much
for those cells ? Could it be they are affected by the FACSFlow ? (would
real PBS be more
advised for such cells during sorting ?)

Any help is welcome.

Best regards,


Fred

Dr Frederic Grosjean
Ludwig Institute for Cancer Research
Lausanne Branch
Chemin des Boveresses 155
1066 Epalinges
Switzerland
Tel. +41-21-692 59 67
Fax. +41-21-653 44 74
www.licr.org








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