Development of a novel artificial red blood cell via membrane emulsification technique

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Construction of three-dimensional tissue in vitro and its implantation are an ultimate goal in tissue engineering. For this purpose, an artificial oxygen carrier is necessary to supply enough oxygen to regenerative tissues. In this research, we fabricated a new oxygen carrier which diameter is between 1 and 5 μm and has a narrow size distribution by the combination of membrane emulsification, non-invasive chemical reaction, and animal cell culture. The oxygen carrier achieved low cytotoxicity of cells by the reduction of endocytosis, high oxygen capacity, and low cost by the use of bovine hemoglobin (Hb). In fact, we achieved all the above-mentioned points. We extracted a fresh and active Hb from bovine blood by 98 % purity, which were confirmed by SDS-PAGE and UV-vis. We successfully emulsified 10 wt% of Hb by preventing the fouling of the membrane by addition of BSA, which emulsion size was ca. 4μm. Subsequent crosslinking by glutaraldehyde brought a new oxygen carrier which P50 was 19mmHg. The toxicity of the carriers to HepG2 was lower than that of the same concentration of Hb.