title: Red blood cell engineering in stroma and serum/plasma-free conditions and long term storage.
abstract: 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. Surprisingly, the viability of erythroid cells was higher in our serum- and feeder-free culture condition than in the previous serum-added condition. This was possible by supplementation with vitamin C in media and hypothermic conditions. 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. This innovative culture technology could contribute to the realization of the large-scale pharming of human RBCs using bioreactor systems.
authors: Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea.
authors: Hyun Ok
authors: HO
authors: Kim
authors: Eun Jung
authors: EJ
authors: Baek
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004907:Erythrocyte Deformability / Q000187:drug effects; D004912:Erythrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D007036:Hypothermia, Induced; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014021:Tissue Preservation
chemical_list: D000975:Antioxidants; D016895:Culture Media, Serum-Free; D001205:Ascorbic Acid