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In vitro generation of artificial red blood cells (RBCs) is very important to overcome insufficient and unsafe blood supply.Despite recent progresses in RBCs engineering from several stem cell sources, none of them could succeed in generation of functional RBCs in the absence of serum/plasma and feeder cells.Without the elimination of serum and plasma, human RBC engineering in a large scale is impossible, especially for the future bioreactor system.Using an appropriate combination of cost-effective and safe reagents, the present study demonstrated the terminal maturation of hematopoietic stem cells into enucleated RBCs, which were functional comparable to donated human RBCs.Also,our report firstly presents the storability of artificial RBCs, which possibility is essential for clinical application.In summary, our report demonstrates engineering of human applicable RBCs with a dramatically enhanced viability and shelf-life in both serum-and stroma-free conditions.