If fresh cancer cells/cell lines derived from malignant ascites are actively proliferating, then one could measure 3H (tritiated)-thymidine incorporation in the presence or absence (control) of the cytotoxic molecule/drug to be tested !

There are plenty of cytotoxicity assays around. You could try Neutral Red Assay, LDH release assay, G6PD release assay or you could stain the cells with PI/Annexin and measure by Flow or microscopy.

Christos Kosmas Thank You Christos for your response but unfortunately we don't have that.

Julie Joseph Hey Julie thanks for your reply. Are you using any I have tried many but in primary culture reproducible of data is very difficult i guess you have experience. So I am facing the challenge to reproduce repeat data using primary culture because you have limited passage number and limited cell number. If you are using any to generate consistent data pls do share.

Thanks

Cytotoxicity can be assessed by Live dead assay using fluorescent probes ( calcein AM and ethidium homodimer-1). This can distinguish live and dead cells accurately. Fluorescent microscopy or flow cytometry can be used for detection of differentially stained cells.

Dr Siddikuzzaman. Yes, I do have experience using many of these assays both on cell lines and on primary cells. If you could tell me which cells you are using and your basic assay setup, I could give you details.

What about labelling the cells with Annexin-V or a fluoresence-labelled anti-epithelial monoclonal antibody (membrane binding)...???

Julie Joseph I am working on Ascies cells and cells derived from disease omentum. In Primari culture in P0 and then P1 we are getting difference in cytotoxicity (IC50) level of same drugs by MTT. Whether SRB will be good idea in simple way I am planning to do SRB hope it will be consistent than MTT.

Thanks