Production of hematopoietic cells from bone marrow endothelial cells 

Hematopoiesis is the process by which all blood cells are produced. In the adult mouse, hematopoiesis occurs predominantly in the bone marrow (BM) where the hematopoietic stem cells (HSC) reside and differentiate into all mature blood cells that are released into the circulation when needed. The HSC, at the top of the hierarchical hematopoietic system, are produced by specialized endothelial cells through a process of endothelial-to-hemogenic transition (EHT) that occurs during mouse development. HSC produced during development sustain hematopoiesis thought adult life. We have recently discovered that endothelial cells in the adult mouse BM can produce small numbers of hematopoietic progenitors and mature blood cells of lymphoid and myeloid lineages. We have also found that these blood cells produced by adult BM endothelial cells are normally functional in the mouse.  These discoveries suggest the possibility that adult endothelial cells in the BM of the mouse, and potentially humans, could be a previously untapped source of hematopoietic cells for transplantation and blood for transfusions. Currently, we do not know how to grow in vitro endothelial cells that can produce large numbers of hematopoietic and blood cells.  In this project, we wish to develop culture conditions that allow hematopoiesis from adult endothelial cells. Key to this project is to try mimic the favorable conditions present in vivo and apply them to cultures of adult endothelium in vitro. Likely key to this project is the development of suitable 3-D culture conditions for endothelial cells that include other cell components and factors present in the hematopoietic BM “niches”. If successful, this would provide a much-needed new source of blood cells for transplantation and other needs. Techniques used will include flow cytometry, FACS sort, mouse bone marrow primary cell isolation, in vitro primary cell culturing, confocal fluorescent microscopy, TA-cloning and long-read sequencing. The student will be paired with a research fellow in the laboratory and will work under supervision to the extent necessary with the goal of making the experience valuable for the student.