Variation of UCB CD34+ in erythroid differentiation potential

Ex vivo manufactured red blood cells (RBC) generated from immortalized erythroid cell lines which can semi-infinitely grow are expected to become a significant alternative in future transfusion therapies. To establish those cell lines, ectopic expression of human papilloma virus (HPV) E6/E7 gene has successfully been employed. In order to induce differentiation and maturation of the immortalized cell lines, terminating the HPV-E6/E7 expression through a gene induction system has been believed to be essential. Here we report that erythroid cell lines established from human bone marrow using simple overexpression of HPV-E6/E7 is capable of normal erythroid differentiation without turning the gene expression off. Newly established cell lines, Erythroid Lines from Lund University (ELLU), are able to differentiate towards mature cells including enucleated reticulocytes upon changing the culture condition but without terminating HPV-E6/E7 gene expression. ELLU cells are heterogeneous, and unexpectedly, clones expressing adult hemoglobin rapidly differentiate but produced cells are fragile while other clones with fetal hemoglobin start expressing adult hemoglobin upon differentiation and give rise to more mature cells. Our findings propose an alternative method to establish immortalized human erythroid cell lines and describe novel cellular characteristics for desired functionally competent clones. 13 healthy donor samples were sequenced in addition to testing their capacity for erythroid differentiation To group A, belong CD34+ cells from human umbilical cord blood, which gave rise to the lower frequency of glycophorin-A+ cells (0-30%) after cultured for 6 days in erythroid differentiation media. To group B, CD34+cells from human umbilical cord blood gave rise to the higher frequency of glycophorin-A+ cells (60-67% ) after being cultured for 6 days in erythroid differentiation media.