Maintenance and in vitro differentiation of mouse embryonic stem cells to form blood vessels.
Embryonic stem (ES) cells, which are derived from developing mouse blastocysts, have the capacity to give rise to all cell types in the adult body. The ability of ES cells to do so has opened the door for novel experimental approaches in the field of developmental biology. Under appropriate culture conditions, ES cells will differentiate and form embryoid bodies (EBs). Upon attachment to a permissive surface, EBs continue a programmed differentiation, and many of the cells differentiated from the EBs reflect those found in the developing embryo and yolk sac, such as hematopoietic cells, endoderm, and endothelial cells. Endothelial cells that arise during ES cell differentiation have the potential to form primitive blood vessels, comparable to the vessels that first form in vivo. This unit describes protocols for maintaining ES cells and the subsequent differentiation of EBs. This unit also provides methods for analyzing vascular marker expression in differentiated ES cultures.
Kappas
Nicholas C
NC
The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Bautch
Victoria L
VL
D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology; D001808:Blood Vessels / Q000166:cytology*; D002417:Cattle; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D018920:Coculture Techniques; D003470:Culture Media; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005312:Fetal Blood; D017930:Genes, Reporter; D007150:Immunohistochemistry / Q000379:methods; D007763:Lac Operon; D051379:Mice; D018919:Neovascularization, Physiologic; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D013194:Staining and Labeling; D016707:Tissue Fixation / Q000379:methods; D001616:beta-Galactosidase / Q000032:analysis
D003470:Culture Media; D001616:beta-Galactosidase