Hello,

        I recently started using Eahy 926 cells(endothelial cells) to study the toxicity and efficacy of liposomal formulations in a 96 well plate (Polystyrene, clear, flat bottom). I add cells to each well (5-10K cells/ well, DMEM medium) and leave it for 12 hours (also tried 36 hours) in an incubator for the cells to attach to the well surface. During aspiration of the media and washing cells with PBS (2 times) using vacuum suction (also tried using a pipette) I observed loss of cells (with 12 hours complete loss in all the wells and with 36 hour incubation partial loss in some wells/ approx 70% loss in some other wells). Even though I treated all the cells under same conditions, (did not let the well go completely dry during aspiration) I observed loss of cells.

       Can anyone please let me know the solutions for this problem, any cautions that I should take while washing cells, and appropriate plate material for endothelial cells.   

I will be grateful for any help you can provide.