Endothelial cells purification - (Dec/28/2006 )
Hi friends, I don't know about you guys, but I must to work for this days, but anyway. My point is that I've been trying to purifie some endothelial cells from aorta sheep, but I have some contamination with fibroblasts. So I just want any sugestion. If someone is working. Thanks. Ahhh and happy new year 
-ATPasa-
QUOTE (ATPasa @ Dec 28 2006, 07:56 AM)
Hi friends, I don't know about you guys, but I must to work for this days, but anyway. My point is that I've been trying to purifie some endothelial cells from aorta sheep, but I have some contamination with fibroblasts. So I just want any sugestion. If someone is working. Thanks. Ahhh and happy new year
I use cloning ring to isolate cells with EC characteristics. You can cut ~0.5 CM from the flat end of an 1 ml pipet tip, spread some grease on the ring and press it to a colony. Then trypsinize it to isolate the cells and grow up from there. Opti-MEM +10% FBS is a good medium for endothelial cells.
-genehunter-1-
Sort them using Di-I-LDL which binds only to endothelials n not fibroblats. If u've alreday done that then resort them..it helps!
-ruchi-