The invention relates to a method for the expansion and differentiation of haematopoietic stem cells into enucleated erythrocytes, in two steps: a first step in a culture medium, where cell proliferation and erythmid differentiation are induced in the presence of growth factors, and a second step modeling a reconstitution of the microenvironment, substantially without erythropoietin (EPO). Optionally, the method of culture may comprise an intermediate step, with haematopoietic growth factors.