Porous membrane device that promotes the differentiation of monocytes into dendritic cells

A method of evaluating the potential reaction of an animal to an agent, said method comprising:

• producing a first well comprising:
• a first porous membrane as the base;
• a ECM material affixed on top of said first porous membrane;
• and
• a second porous membrane affixed on top of said ECM material;
• inverting said first well into a second well comprising cell media;
• culturing endothelial cells on bottom of said first porous membrane;
• applying peripheral blood mononuclear cells (PBMCs) to the endothelial cells;
• after ˜1.5 hours washing said PBMCs and said endothelial cells off of the bottom of said first porous membrane, wherein dendritic cells are now present in said ECM material;
• removing said first well from said second well comprising cell media and placing said first well with said second porous membrane facing up into a third well comprising as its base a three-dimensional artificial lymphoid tissue, comprising a second ECM material and a plurality of lymphocytes and leukocytes;
• applying an agent to the top of said second porous membrane, said antigen allowing the dendritic cells to migrate out of said first ECM material and into said three-dimensional artificial lymphoid tissue;
• and
• evaluating the immune response to said agent.

A method for generating large numbers of dendritic cells comprising:

• producing a first well comprising:
• a first porous membrane as the base;
• endothelial cells cultured on the bottom of said first porous membrane;
• a second porous membrane situated above, and separated from, said first porous membrane;
• endothelial cells cultured on the top of said second porous membrane;
• and
• cell culture media comprising an agent located between said first porous membrane and said second porous membrane;
• inverting said first well into a second well comprising cell media;
• applying peripheral blood mononuclear cells (PBMCs) to the endothelial cells cultured on the top of said second porous membrane;
• at least about 48 hours after application of the PBMCs, removing said first well from said second well;
• and
• isolating dendritic cells from said second well.