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ELISpot assays with adherent stimulator cells: is it possible?

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Posted 29 January 2015 - 07:15 AM

Hello Bioforumers,


I am trying to compare the immunogenicity of several types of cells in order to predict their likelihood of being rejected when transplanted into an animal host. After a fair bit of reading up and weighing all the pros and cons of different techniques, I have decided to use ELISpot assays with human PBMCs as responder cells. However, my stimulator cells are adherent cells. All the protocols I have seen so far involve stimulator cells that are non-adherent, mainly PBMCs or T-cells from another individual. 


Does anyone have any experience doing ELISpot assays with adherent stimulator cells? I imagine that when the stimulator cells adhere to the ELISpot membrane, they would present a physical barrier and block factors secreted by the responder cells from reaching the membrane and thus interfering with spot formation. I would appreciate it greatly if anyone could share some experience or knowledge about this.





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