> 1) We want to stimulate whole blood for up > to 24 hours and analyse the monocytes afterwards. > In which sort of tubes should we keep the blood > or is it possible to block the attachment of the > monocytes to the tube material? we use polypropylene tubes (aka. layering tubes, 12 ml, with loose caps). My thoughts on the attachement part are unclear, see answ. to 3) > 2) If the monocytes are attached to the wall, is it > possible to include them in an analysis or are all > of them stimulated by the attachment per se? My understanding is: monocytes get activated THEN they attach. I guess that is a part of the macrophage migration from periphery to tissue. > 3) We see that one have to predilute the blood in > RPMI or AIM-V medium before one stimulate the > cells. Which medium is the better one and why > do one have to dilute it? the cells will run out of "food" stored in whole blood. We use L-Glu/PenStrep RPMI (L/Glu added right beforehand). Interestingly my observations are such: 1. unstimulated monocytes in whole blood will disappear (no cells CD14+ or CD33+) (24hrs) 2. unstimulated monocytes in whole blood, diluted 1:1, are unchanged from regular whole blood. 3. stimulated monocytes in whole blood will not disappear 4. stimulated monocytes in diluted whole blood will not disappear, but the activation will be LESS than in the undiluted sample. > 4) Which heparin blood is the better one, Li-heparin or Na-heparin? > I see the question from yesterday and hope > to get the answer as well. We only use Sodium. I think Lithium forms a larger ion in solution and creates problems with activation profiles. Probably competes with Calcium or something. I never looked into IL-18 availability, sorry. Regards and good luck, Maciej Simm __________________________________________________ Do You Yahoo!? Yahoo! Auctions - buy the things you want at great prices http://auctions.yahoo.com/
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