Isolation and Culture of Endothelial Cells from the Embryonic Forebrain

摘要: Embryonic brain endothelial cells can serve as an important tool in the study of angiogenesis and neurovascular development interactions. The two vascular networks embryonic forebrain, pial periventricular, are spatially distinctive have different origins growth patterns. Endothelial from periventricular unique gene expression profiles functions. Here we present a step-by-step protocol for isolation, culture, verification pure populations network (PVECs) forebrain (telencephalon). In this approach, telencephalon devoid membrane obtained day 15 mice is minced, digested with collagenase/dispase, dispersed mechanically into single cell suspension. PVECs purified suspension using positive selection anti-CD-31/PECAM-1 antibody conjugated to MicroBeads strong magnetic separation method. Purified cultured on collagen 1 coated culture dishes medium until they become confluent further subcultured. exhibit cobblestone spindle shaped phenotypes, visualized by phase-contrast light microscopy fluorescence microscopy. Purity PVEC cultures was established markers. our hands, method reliably consistently yields PVECs. This will benefit studies aimed at gaining mechanistic insights angiogenesis, understanding interactions, cross-talks neuronal types holds tremendous potential therapeutic angiogenesis.