Isolation and Culture of Human Hematopoietic Progenitors for Studies of Dendritic Cell Biology

Understanding the regulation of distinct dendritic cell (DC) function and differentiation pathways is important in many physiological and pathophysiological processes. This includes infectious and neoplastic diseases, vaccination and immunotherapy, allograft rejection, and the pathogenesis of autoimmune diseases. Isolation and culture of human hematopoietic progenitor cells provide a valuable model for studies on DC biology and may help uncover new means to manipulate DC differentiation and function in therapeutic settings. Here, a detailed protocol for the isolation of CD34⁺ hematopoietic progenitor cells from human cord blood is described. The isolated cell population consists of approximately 85% CD34⁺ CD45⁺ hematopoietic progenitor cells that in response to granulocyte-macrophage colony-stimulating factor (GM-CSF) plus tumor necrosis factor (TNF) expand and differentiate into CD11c⁺ HLA-DR⁺ DC-expressing CD1a.