FIG. 2.

In vitro culture of LN stroma produces a pure LN LEC population. Flow cytometry analysis of freshly dissociated mouse LN stroma and of P8 and P22 cultures, stained with PE-Cy7-anti-CD45, APC-anti-podoplanin, and PE-anti-CD31 antibodies. Forward scatter (FSC) and side scatter (SSC) demonstrate large size of cells in P8 and P22 cultures. All P8 and P22 cells gated on this large population are CD45-ve CD31+ podoplanin+, relative to mixed composition of freshly dissociated LN stroma stained for podoplanin and CD31, which contain LEC, BEC, FRC, and DN stromal cells. Two independent cell isolates were analyzed.