These cells were isolated from lung cancer xenograph. I have been culturing them in DMEM/F12/B27/FGF for over a month now. They looked like fibroblasts to me but they are forming colonies.
Thank you for your inputs.
Hi,
I think it can be adenocarcinoma, which means epithelial origin. Clustering can mean the presence of "cancer stem cells", which self-renew and generate other cancer cell types, more differentiated. Try to stain for epithelial markers, such as E-cadherin and keratin.
Hi,
The first reaction I thought these cells are endothelial but they can not grow in DMEM. It is possible that they are fibroblast as you mentioned but have sorry to say (Andrey) that they don't look as epithelial cells for me. The easiest way is to run IHC against different receptors such EGFR or FGFR or even VEGFR (in case these cells are modified endothelial cells).
Good Luck
Hi Mathieu,
if I have well-understood the experiment, this cells are coming from tumour tissue of a mouse xenograft model. Therefore, I assume you injected human cells in a mouse. Was this the procedure used ? Did you inject a cell line or a patient-derived culture?
If this is the case, you run a qPCR on you cells for human actin and mouse actin, you should see signal only (or predominantly) from human PCR. You didn't inject fibroblasts or endothelial cells, therefore they should be positive only to mouse actin. In theory this is applicable to any specific qPCR for mouse or human genes.
If your PCR is positive for human, this will tell you that you have a quite well-enriched population of cancer cells. Then, I agree with with Fuad and Andrey, you should further prove it by epithelial markers staining (keratins).
Good luck
Ivano
Dear all,
I agree with everybody on the need to stain these cells with cell specific markers.
Ivano: I did not prepare the xenograph but I know that these cells were from a lung adenocarcinoma patient resection sample.
Thank you for your time and inputs.
Mathieu
Unfortunately we are still using 50 yo technology to diagnose the origin, in 2015, of hyperactive cell lines. That's what we get for letting are most creative scientist get away and build rockets and assorted modern technology in other fields.
Hi Matt
Nice to see you on ResearchGate. I think the first thing to do is make a cell block and stain for pan-cytokeratins. You can use a cocktail like MNF116 or AE1/AE3, which include low and high molecular weight cytokeratins. These are routinely used in diagnostic histopathology labs and are well optimised. I would actually try 2 cytokeratin markers and make sure they are both negative as sometimes carcinomas (especially spindle cell carcinomas) can have weak patchy keratin positivity and can lose some keratins so it's worth having 2 cocktails come out as negative before labelling the cells as non-epithelial. Good luck.
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