mouse splenocyte preparation
Posted 16 October 2000 - 09:00 PM
Posted 14 November 2000 - 10:00 PM
Posted 21 November 2000 - 10:00 PM
Posted 18 October 2001 - 09:00 PM
Posted 09 September 2004 - 05:27 AM
2.12.3 Preparation of Spleen Cells
Adult murine spleen cells were used for these applications. The spleens were removed and collected in a sterile wire sieve over a Petri dish half filled with media. The spleens were gently pressed through the sieve using a rubber policeman (which is the plunger in a syringe, usually a 3-5 ml syringer plunger is great), all the while gently applying fresh media on the spleen to keep the cells moist and add transfer through the sieve. The sieve was rinsed over the Petri dish with additional media to remove all remaining cells. The disaggregated cells were transferred to a fresh 30 ml tube (Filtrona) and centrifuged at 2,000 rpm for 5 mins (any tube size will do, we used Filtronas because they were the ideal size for our contrifuge). The cell pellet was resuspended in 10 ml aMEM + 10 % FBS (or which media you are planning to use) and counted.
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Posted 11 December 2009 - 02:25 PM
Posted 21 February 2010 - 08:40 PM
I use to isolate mouse splenocyte by a very simple way.1/ Cut mouse spleen into several small pices. Then gine spleen pices in metal mesh cup (if you done have one, you can grine spleen bettwen 2 microscope glass slides). 2/Remove all big connective tissue and debris. Collect cell suspension in 15ml facoll tube anf leave the tube stand up about 3-5 mine. The remain debris will come down to the bottom oh the tube.3/ Collect cell suspension to other tube, erythocyte hemolysis, and use for cell proliferation test.<p>By 3/
Hi, Does anybody use lipopeptide antigens in immunological reactions? What tests are best suited for them? Can we use MQ water to reconstitute synthetically prepared lipopetides? What about culture medium?