Isolation and Culture of Mouse Placental Endothelial Cells
Isolation and culture of endothelial cells (ECs) is a useful tool to study the cellular processes involved in vascular development and vascular maturation. In this chapter, we describe a method to isolate and culture endothelial cells from placentae. This method takes advantage of two transgenes: ROSA26 mT/mG , which drives the expression of GFP upon Cre-mediated recombination, and Tie2-Cre, which expresses Cre driven by the Tie2 promoter in endothelial progenitors and their descendants. GFP-expressing endothelial cells are isolated through fluorescence-activated cell sorting (FACS). The sorted cells express the endothelial marker CD31. This method can be used to study the morphological and physiological properties of placental endothelial cells in mice carrying mutations affecting vascular development.
Key wordsMouse placenta Endothelial cells Fluorescence-activated cell sorting Primary cell culture
We thank The SickKids-UHN Flow Cytometry Facility for help with FACS, and The Centre for Phenogenomics (TCP) for mouse husbandry. This work was supported by the Heart and Stroke Foundation of Canada (G-17-0018613), the Natural Sciences and Engineering Research Council of Canada (NSERC) (500865), the Canadian Institutes of Health Research (CIHR) (PJT-149046), and Operational Funds from the Hospital for Sick Children to P.D.-O.
- 6.Dong QG, Bernasconi S, Lostaglio S, De Calmanovici RW, Martin-Padura I, Breviario F, Garlanda C, Ramponi S, Mantovani A, Vecchi A (1997) A general strategy for isolation of endothelial cells from murine tissues. Characterization of two endothelial cell lines from the murine lung and subcutaneous sponge implants. Arterioscler Thromb Vasc Biol 17(8):1599–1604CrossRefPubMedGoogle Scholar