Hello everyone,
I am cultivating patient derived organoids from pancreatic cancer patients and I saw at the bottom of my domes some "strange" cells other than my usual organoids. I know I should perform some IHC to understand the identity of these cells but I was wondering if some of you could help me to have a general idea of their possible origin. They don't seem fibroblasts to me, so maybe are they normal ductal pancreati cells?
Thanks in advance
What is the shape and size of these cells? Elongated, cuboidal, or round compared to your usual organoids?
Hello Dinah Lilia Mourise
These cells are very different from my organoids. First, they are adherent, but my organoids are round and growing in a 3D shape inside matrigel domes...
Hi Giulia,
What I would do is to first examine the cells under different magnifications for shape, size, and other features. Then you want to compare them (epithelial cells, fibroblasts, and immune cells).
Then, IHC for markers like CK19, Vimentin, CD31, and CD45 to identify the cell types. I would analyze gene expression by keeping in mind the bulk or single-cell RNA-seq for comprehensive profiling.
Wait for any growth rates and migration properties. Any difference?
Have you checked if the media or Matrigel might favor different cell populations?
Its normal for the sample to have various cell types, including cancer-associated fibroblasts or normal pancreatic cells.
I would isolate these cells for separate study.
Use the approach of combining microscopy, IHC, molecular analysis, and functional assays. It'll help to see the cells and understand their origin.
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