Creating an in-vitro co-culture model of Human Megakaryocytic-Erythroid Progenitors and Endothelial Cells

Previous studies have suggested that endothelial cells promote red blood cell (RBC) production in human bone marrow, which has implications for improved treatments for anemia, cancer, and reproduction. However, investigation of this phenomenon is hindered by a dearth of studies that have successfully isolated primary endothelial cells from human bone marrow. Using flow cytometry, magnetic cell separation, and primary cell culture techniques we tested the hypothesis that adult bone marrow-derived endothelial cells can be isolated from previously frozen bone marrow samples by selecting for CD144+ expressing cells and culturing them in vitro. Magnetic cell separation utilizing the marker CD144 to isolate adult bone marrow-derived endothelial cells was not effective from human bone marrow samples that had already been depleted of CD34+ cells. In future experiments we will test the ability to magnetically separate CD144+ adult bone marrow derived endothelial cells from freshly collected bone marrow prior to CD34+ selection. Results of these studies will generate a model that we can use in the future to test the molecular mechanisms by which human bone marrow-derived endothelial cells promote erythroid differentiation