Neutrophils activation- HELP

[Mailendj]

Hi!!!
I have a question....what's the best way to transport whole blood samples from farm (dairy cattle's breeding) to my laboratory??? I usually use a thermic bag.... but I don't know if different solution are necessary to conserve neutrophils vitality!!! Now here it's winter... and cells recovery and vitality are low!!!
Thank's...

Milena

[Warthaug]

Mailendj, a couple Q's before I try to answer your question:

1) How long is the period of time between blood collection and when you start your isolation?
2) How cool does the blood get during transport?
3) What anti-coagulant are you using?
4) Have you successfully isolated neutrophils from this same source in the past?

Bryan

[Mailendj]

Answers:
1) 5 min of travel by car
2) I put tubes into a termic bag... so blood's temperature decrease slowly... now here it's 4°C.
3) EDTA
4) yes, but not in this season!! Isolations made in spring and summer (max 28°C) were good!!

Tell me if I'm not clear...

[Warthaug]

Originally Posted by Mailendj

Answers:
1) 5 min of travel by car
2) I put tubes into a termic bag... so blood's temperature decrease slowly... now here it's 4°C.
3) EDTA
4) yes, but not in this season!! Isolations made in spring and summer (max 28°C) were good!!

Tell me if I'm not clear...

You should not be having that many problems with only 5min between the draw and getting to the lab. Letting the blood cool shouldn't be too much of a issue - we always do our preps with room-temp blood and it works fine. My gut feeling is the EGTA may be a problem; its quite hard on neutrophils, which is why I prefer citrate or heripin. None-the-less, this was working earlier, which suggest the issue is elsewhere. Have you recently re-made some of your isolation reagents? Perhaps one is off.

Also, what method are you using for your preps?

Bryan

[Hermosaorb]

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