Macrophages + Vitamin A

We are interested in whether peritoneal LCM are programmed by omental Wt1+ stroma (identified as PDPN+ cells). To this end, we co-cultured omental PDPN+ cells (purified by MACS by depleting CD45+ and CD31+ cells) with peritoneal exudates in a manner that enables cell:cell contact or inhibited this with a transwell insert or enabled cell:cell contact in the presence of the retinoic acid receptor inverse agonist BMS493. Peritoneal exudates were also cultured alone or with retinoic acid (1uM). After 5-7 days PDPN+ cells and F480+ cells were FACS sorted from cultures directly into RLT Buffer. Ex vivo peritoneal LCM were also FACS-sorted for comparison. PRJ0019587 Overall design: In vitro samples were FACS sorted as either F480+ or PDPN+ directly into RLT Buffer.