title: Iron dose-dependent differentiation and enucleation of human erythroblasts in serum-free medium.
abstract: Improvements in ex vivo generation of enucleated red blood cells are being sought for erythroid biology research, toward the ultimate goal of erythrocyte engineering for clinical use. Based upon the high levels of iron-saturated transferrin in plasma serum, it was hypothesized that terminal differentiation in serum-free media may be highly dependent on the concentration of iron. Here adult human CD34(+) cells were cultured in a serum-free medium containing dosed levels of iron-saturated transferrin (holo-Tf, 0.1-1.0 mg/ml). Iron in the culture medium was reduced, but not depleted, with erythroblast differentiation into haemoglobinized cells. At the lowest holo-Tf dose (0.1 mg/ml), terminal differentiation was significantly reduced and the majority of the cells underwent apoptotic death. Cell survival, differentiation and enucleation were enhanced as the holo-Tf dose increased. These data suggest that adequate holo-Tf dosing is critical for terminal differentiation and enucleation of human erythroblasts generated ex vivo in serum-free culture conditions. Published 2013. This article is a US Government work and is in the public domain in the USA.
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Colleen
authors.initials: C
authors.lastname: Byrnes
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Y Terry
authors.initials: YT
authors.lastname: Lee
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.|Center for Cancer and Blood Disorders, Children's National Medical Center, Washington, DC, USA.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Meier
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Antoinette
authors.initials: A
authors.lastname: Rabel
authors.affiliation: Clinical Chemistry Service, Department of Laboratory Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD, USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Sacks
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jeffery L
authors.initials: JL
authors.lastname: Miller
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free; D004900:Erythroblasts / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007501:Iron / Q000494:pharmacology*; D014168:Transferrin / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D014168:Transferrin; D007501:Iron
keywords: enucleation; erythropoiesis; haemoglobin; holo-transferrin; iron; serum-free media