I would like to measure the inflammatory response triggered by human stem-cell derived neurons with/without focal delivery of anti-inflammatory molecule. To do it, I'd like to establish a co-culture of neurons and PBMC and measure the immune cells proliferation and level of cytokines (3-5 days after co-culture) with or without the drug. However, I am not sure about the effect of neuronal media on immune cells, the ratio of the different cell types and how to set up the media changes. I usually change the media every 48h for the neurons but I need to avoid it here, since I don't want to aspirate the immune cells or the cytokines. Does anyone have experience with that?
Dear Chiara!
Please You look at the folowing article:
Article Cross-Talk between Human Neural Stem/Progenitor Cells and Pe...
Human NSC and immune cell co-cultures
PBMCs were collected from normal adult human peripheral blood (n = 9) with oral consent based on HHS regulations under (45 CFR 46.116(c) or (d)), which was approved by ethics committees of the First Affiliated Hospital. PBMCs isolated by density gradient centrifugation under sterilized conditions. Collected neurospheres were cultured onto PLL-coated cover glasses in 24-well plates at a density of 1.5×106 cells/well in 1 ml of RPMI-1640 complete medium with 5% FBS until cells grew to 60–70% confluence. The ratio of hNSCs:PBMCs in detection of lymphocyte subtype was 1:10, 1:1 and 5:1 respectively and then at a ratio of 5:1 in other co-culture groups. The PBMCs were then loaded for the co-culture with the plated hhNSCs for 2 days. Cultured PBMCs- or hNSPCshNSCs-only were designated as control groups, respectively.
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