Ferric citrate and apo-transferrin enable erythroblast maturation with β-globin from hemogenic endothelium

• Type of Material: Article
• Authors: Jong-Hee Lee, Korea University of Science and Technology; Ji-Hoon Lee, CHA University; Soo-Been Jeon, CHA University; Hyebin Koy, Korea Research Institute of Bioscience and Biotechnology; A-Reum Han, CHA University; Jieun Kim, Korea Research Institute of Bioscience and Biotechnology; Sunghun Lee, CHA University; Jae-Ho Lee, Keimyung University; Seung-Soon Im, Keimyung University; Young-sup Yoon, Emory University
• Language: English
• Date: 2023-08-25
• Publisher: Springer Nature
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2023
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): https://doi.org/10.1038%2Fs41536-023-00320-4
• Title of Journal or Parent Work: NPJ Regenerative Medicine
• Volume: 8
• Start Page: 46
• Grant/Funding Information: This work was supported by National Research Foundation of Korea (NRF) funded by the Korea government (MSIT) (No. 2021R1A2C1004571) and (NRF-2019R1A2C1086988), and KRIBB Research Initiative Program Grant (KGM4562323).
• Keywords: cell therapy; Red blood cell (RBC) generation; Embryonic stem cells; regenerative medicine; Cell growth
• Research Categories: Biology, Cell

Abstract

Red blood cell (RBC) generation from human pluripotent stem cells (PSCs) offers potential for innovative cell therapy in regenerative medicine as well as developmental studies. Ex vivo erythropoiesis from PSCs is currently limited by the low efficiency of functional RBCs with β-globin expression in culture systems. During induction of β-globin expression, the absence of a physiological microenvironment, such as a bone marrow niche, may impair cell maturation and lineage specification. Here, we describe a simple and reproducible culture system that can be used to generate erythroblasts with β-globin expression. We prepared a two-dimensional defined culture with ferric citrate treatment based on definitive hemogenic endothelium (HE). Floating erythroblasts derived from HE cells were primarily CD45+CD71+CD235a+ cells, and their number increased remarkably upon Fe treatment. Upon maturation, the erythroblasts cultured in the presence of ferric citrate showed high transcriptional levels of β-globin and enrichment of genes associated with heme synthesis and cell cycle regulation, indicating functionality. The rapid maturation of these erythroblasts into RBCs was observed when injected in vivo, suggesting the development of RBCs that were ready to grow. Hence, induction of β-globin expression may be explained by the effects of ferric citrate that promote cell maturation by binding with soluble transferrin and entering the cells. Taken together, upon treatment with Fe, erythroblasts showed advanced maturity with a high transcription of β-globin. These findings can help devise a stable protocol for the generation of clinically applicable RBCs.