I have a problem with the endothelial cell isolation from dog corpus luteum. When I isolate them, they are forming tube like formation in non-coated flask. They are not stimulated either. However, in other species same cells are showing cobble stone structure. I put picture in attachment. Is this normal, have you ever seen this kind of formation ?
Those are not tube like. Those are spindle like. Probably fibroblast. You are getting only cell contamination. You should improve your isolation protocol.
I agree with Ruben, those cells look more like fibroblasts than EC tubes.
Try staining the cells for EC markers, like CD146, CD31, CD144.
Hi Aykut,
Your cells do not look like endothelial and probably, as said before, you might have contamination with other cells. Extraction of endothelial cells from any source is a complex task and I had problems extracting endothelial cells from the mouse aorta. They are very demanding, needing special media and coating. I am attaching one pdf file that might help you. It is a review article and you can find some info about microvascular endothelial cells
isolation from the corpora lutea of different species and their maintenance.
Good luck.
Milica
Hi,
They doesn't look like ECs, they may be fibroblast and smooth muscles cells. As you're not explained here which method you're using to isolate i can suggest you to try using magnetic beads coated with either BS1 or CDs and check with battery of genes related to endothelial cells as well as other major contaminates such as fibroblast and smooth muscles cells. you can use the attached protocol which i'm using for bovine CL.
All The best
Raghu
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