Tumor cell isolation from ascites and culture phenotype

[Biologos]

Dear all,

i have recently started working in Ovarian cancer. I usually isolate Tumor cells and PBMC from ascitic fluid with the use of Ficoll. Does anyone know how to separate PBMC from tumor cells without the use of magnetic beads?

I was thinking of maybe plating cells after Ficoll isolation straight and after 16h or so remove the supernatant that contains PBMCs. Has anyone done this? Any sucess? How long will it take for the tumor cells to "lose" their phenotype? I guess that after 16h they primary isolated cells should retain their phenotype.

Also which marker would you suggest in order to screen for phenotypic change?

Many thanks!