Abstract 2096

We have formulated a robust culture system simulating human erythropoiesis, producing mature reticulocytes from adult peripheral blood CD34+ progenitor cells in 20 days. Expansion of cell numbers over the course of the culture exceeded 104-fold and enucleation efficiency was up to 90% without the use of feeder layers. Mature reticulocytes and erythrocytes were isolated from culture by passage through a standard leucocyte filter. Scanning and transmission electron microscopy of the reticulocytes revealed morphological features indistinguishable from those of mature reticulocytes isolated from the peripheral blood of normal donors. Post-filtration cells expressed adult haemoglobin and had a capacity to bind and release oxygen comparable to that of adult peripheral blood erythrocytes. Examination of blood group active- and other cell surface antigens revealed a normal glycosylation profile and normal cell surface protein expression. Most notably, the cells did not express cryptantigens T and Tn. LORCA analysis indicated that cultured cells had slightly reduced membrane stability and deformability compared to peripheral blood erythrocytes. Haematology parameters demonstrated a typical mean cell volume of 131fL and mean cell haemoglobin of 37.7pg. This, scalable approach has allowed us to culture cells from a single apheresis cone to produce 1010 reticulocytes, which represents a packed cell volume of 1ml. These results clearly demonstrate the feasibility of producing cultured red cell products suitable for transfusion therapy ex vivo from adult peripheral blood stem cells.


No relevant conflicts of interest to declare.

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