Isolation of murine bone marrow scavenging sinusoidal endothelial cells.
The bone marrow (BM) is permeated with sinusoidal vessels lined with sinusoidal endothelial cells (SEC), which are crucial for BM physiology and hemopoietic stem cell (HSC) renewal. However, little is known about the characteristics or functional capacity of bone marrow sinusoidal endothelial cells (BMSEC). One significant barrier to the study of BMSEC is the lack of specific cell surface markers that can be used to isolate these cells. Nevertheless, BMSEC possess one exceptional property, namely, the ability to scavenge large amounts of soluble waste molecules such as advanced glycation end-products (AGE) and we have utilized this to label BMSEC for cell sorting and isolation. We describe the means to produce and fluorescently label AGE, its use as a functional in vivo marker of BMSEC and the isolation of these cells from murine BM using fluorescent activated cell separation (FACS).
Department Medical Biology, University of Tromsø, Tromsø, Norway.
Peter A G
PA
McCourt
Ana
A
Oteiza
Benjamin
B
Cao
Susan K
SK
Nilsson
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D017127:Glycation End Products, Advanced / Q000032:analysis; D018189:Immunomagnetic Separation / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D005456:Fluorescent Dyes; D017127:Glycation End Products, Advanced; D016650:Fluorescein-5-isothiocyanate