I am trying to stain some cardiac fibroblasts (isolated from neonatal rat hearts) to look at effects of fibrosis from gene knockdowns. Does anyone know of any protocols like this? Everything I have found so far only uses tissue samples.
Hi Caitlin,
I did use Picosirius Red staining for orientation of collagen fibres. Please find my reprint and see the Figure 8. This paper is not in Pubmed. Once I get some time, I can try to send some other literature on this issue that will clarify your basic knowledge. After doing red stainng, you must see the cells or slides under polarizd light that will be additional fun. Good luck
You may culture cells on larger cover slips in a plate and than stain the cells while attached to cover slip. I am sure that will work, some cells may fall during procedure but you should get some cells to watch
Just be aware that this will stain ECM. In vivo, the ECM is organized and a 2D cell culture model will not sufficiently represent this condition. A more valuable method would be to analyze ECM proteins in supernatants/protein lysates or stain directly for markers of fibroblast activation (e.g. aSMA).
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