I have been working on peripheral blood lymphocytes (PBL) of cancer patients. It shows a high rate of spontaneous genomic instability is in progress. When I checked NDI, the PHA induced PBL of cancer patients showed an increased NDI compared to normal healthy subjects. My question is: can 'lymphocytes', of solid tumor/cancer patients, proliferate in high rate? If so, how do we interpret NDI?
Dear Alex,
PBL of Cancer patients is already in proliferating stage. That is why we should not add PHA to stimulate PBL of cancer patients. Please try to culture without PHA and go for 24 hr and 48 hr incubation. I am sure you will get appropriate NDI for your case and control.
Thank you so much sir, but if i need to compare those results with the control PBL what method i should use.? When we do CBMN Cyt assay, the culture initiation with PHA and cytokinesis block using Cyto-B are necessary !! can we do CBMN assay with out PHA stimulation ?!
Alex, In CBMN assay we need not to look for chromosomes, so we can do the test without using PHA also. It does not matter. If you are compaing the cases with the controls then you need not to use PHA although you can harvest control sample in 24 hr or 48 hr also. You can score 20 metaphases which is enough to do statistical analysis.
sir, i'm not looking for the chromosomes or metaphases!! I'm performing CBMN assay to find out nuclear anomalies and NDI.. so it's not possible to harvest the culture @ 24/48th hour!! just want to know if we get an NDI value how we'll interpret it!
Alex, one suggestion! Keeping aside CBMN assay for cancerous patients, if you go for conventional cytogenetics you will get outstanding results.
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