Description

Cell differentiation along the erythroid lineage arises from hematopoietic stem cells within the bone marrow. The cell surface marker CD36, which is considered an early marker of erythrocyte differentiation, is acquired as the differentiation of this lineage progresses. However, the rarity and heterogeneity of the CD36 population makes it difficult to isolate in vivo. In order to obtain CD36+ erythroid progenitor cells, CD34+ hematopoietic stem cells are isolated and cultured in serum-free expansion media supplemented with factors formulated to selectively promote the expansion and differentiation of human erythroid progenitor cells.

Human CD36+ erythroid progenitor cells are derived from cultured bone marrow CD34+ stem cells. Bone marrow mononuclear cells (MNCs) are separated from whole bone marrow by a density gradient centrifugation protocol. CD34+ stem cells are selected using immunomagnetic anti-CD34 microbeads from the MNC population. The isolated CD34+ cells are cultured in StemSpan™ SFEM, a serum-free expansion media for hematopoietic cells, supplemented with SCF, EPO and IL-3. Cultured cells are harvested after 10 days and CD36+ cells and are characterized by flow cytometry before cryopreservation to ensure a highly pure and viable cell population.

Cells were obtained using Institutional Review Board (IRB) approved consent forms and protocols.