I have been trying to isolate human adipose microvascular endothelial cells from human omental and subcutaneous fat. I use CD34 and CD31 immunoseperation techniques for the cells to isolate out CD34+31+ double positive cells into culture. When I am initially growing the culture, they have a cobble-stone morphology and very endothelial-like, but when I passage them into larger flasks these cells will loose their morphology and become spindle and elongated and when I run these samples for flow cytometry for CD34 and CD31 they have no expression of the markers. I believe my cells are de-differentiating in culture and losing their markers.
What medium? Are you using a microvascular growth supplement? Treatment on the flask?
Hi Bronson, can you please give more explanation of adipose/endothelial cells?
I have never heard of them before.
Hi Dr. Reiserer,
We use a Lonza Media Kit for growing microvascular endothelial cells that do contain growth supplements. We also use fibronectin to coat the flasks.
Hi Stefansson,
We are attempting to isolate microvascular endothelial cells from human fat using a double positive immunoselection protocol from published manufacturers. These cells should be double positive for CD34 and CD31 when employing the protocol.
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology