Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea.
Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea.
Stem Cell Res Ther. 2023 Apr 26;14(1):106. doi: 10.1186/s13287-023-03305-8.
Blood transfusion is an essential part of medicine. However, many countries have been facing a national blood crisis. To address this ongoing blood shortage issue, there have been efforts to generate red blood cells (RBCs) in vitro, especially from human-induced pluripotent stem cells (hiPSCs). However, the best source of hiPSCs for this purpose is yet to be determined.
In this study, hiPSCs were established from three different hematopoietic stem cell sources-peripheral blood (PB), cord blood (CB) and bone marrow (BM) aspirates (n = 3 for each source)-using episomal reprogramming vectors and differentiated into functional RBCs. Various time-course studies including immunofluorescence assay, quantitative real-time PCR, flow cytometry, karyotyping, morphological analysis, oxygen binding capacity analysis, and RNA sequencing were performed to examine and compare the characteristics of hiPSCs and hiPSC-differentiated erythroid cells.
hiPSC lines were established from each of the three sources and were found to be pluripotent and have comparable characteristics. All hiPSCs differentiated into erythroid cells, but there were discrepancies in differentiation and maturation efficiencies: CB-derived hiPSCs matured into erythroid cells the fastest while PB-derived hiPSCs required a longer time for maturation but showed the highest degree of reproducibility. BM-derived hiPSCs gave rise to diverse types of cells and exhibited poor differentiation efficiency. Nonetheless, erythroid cells differentiated from all hiPSC lines mainly expressed fetal and/or embryonic hemoglobin, indicating that primitive erythropoiesis occurred. Their oxygen equilibrium curves were all left-shifted.
Collectively, both PB- and CB-derived hiPSCs were favorably reliable sources for the clinical production of RBCs in vitro, despite several challenges that need to be overcome. However, owing to the limited availability and the large amount of CB required to produce hiPSCs, and the results of this study, the advantages of using PB-derived hiPSCs for RBC production in vitro may outweigh those of using CB-derived hiPSCs. We believe that our findings will facilitate the selection of optimal hiPSC lines for RBC production in vitro in the near future.
输血是医学的重要组成部分。然而，许多国家都面临着全国性的血液危机。为了解决这一持续的血液短缺问题，人们一直在努力在体外生成红细胞（RBC），特别是从人诱导多能干细胞（hiPSC）中生成。然而，用于此目的的最佳 hiPSC 来源尚未确定。
在这项研究中，使用 episomal 重编程载体从三种不同的造血干细胞来源（外周血[PB]、脐带血[CB]和骨髓抽吸物[BM]）建立了 hiPSC（每种来源 3 个样本），并将其分化为功能性 RBC。进行了各种时程研究，包括免疫荧光分析、定量实时 PCR、流式细胞术、核型分析、形态分析、氧结合能力分析和 RNA 测序，以检查和比较 hiPSC 和 hiPSC 分化的红细胞的特征。
从这三种来源都建立了 hiPSC 系，发现它们具有多能性且具有相似的特征。所有 hiPSC 都分化为红细胞，但分化和成熟效率存在差异：CB 来源的 hiPSC 最快成熟为红细胞，而 PB 来源的 hiPSC 成熟需要更长的时间，但具有最高的可重复性。BM 来源的 hiPSC 产生了多种类型的细胞，分化效率较差。尽管如此，来自所有 hiPSC 系的红细胞主要表达胎儿和/或胚胎血红蛋白，表明发生了原始红细胞生成。它们的氧平衡曲线都向左偏移。
总的来说，尽管存在需要克服的一些挑战，但 PB 和 CB 来源的 hiPSC 都是体外生成 RBC 的可靠来源。然而，由于 CB 来源的 hiPSC 数量有限且所需 CB 量较大，以及本研究的结果，使用 PB 来源的 hiPSC 进行体外 RBC 生成的优势可能超过使用 CB 来源的 hiPSC。我们相信，我们的研究结果将有助于在不久的将来选择用于体外 RBC 生成的最佳 hiPSC 系。
