Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, 27 Taiping Road, Beijing 100850, China.
Biomed Res Int. 2013;2013:807863. doi: 10.1155/2013/807863. Epub 2013 Jan 30.
In vitro models of human erythropoiesis are useful in studying the mechanisms of erythroid differentiation in normal and pathological conditions. Here we describe an erythroid liquid culture system starting from cord blood derived hematopoietic stem cells (HSCs). HSCs were cultured for more than 50 days in erythroid differentiation conditions and resulted in a more than 10(9)-fold expansion within 50 days under optimal conditions. Homogeneous erythroid cells were characterized by cell morphology, flow cytometry, and hematopoietic colony assays. Furthermore, terminal erythroid maturation was improved by cosculturing with human fetal liver stromal cells. Cocultured erythroid cells underwent multiple maturation events, including decrease in size, increase in glycophorin A expression, and nuclear condensation. This process resulted in extrusion of the pycnotic nuclei in up to 80% of the cells. Importantly, they possessed the capacity to express the adult definitive β -globin chain upon further maturation. We also show that the oxygen equilibrium curves of the cord blood-differentiated red blood cells (RBCs) are comparable to normal RBCs. The large number and purity of erythroid cells and RBCs produced from cord blood make this method useful for fundamental research in erythroid development, and they also provide a basis for future production of available RBCs for transfusion.
体外人红细胞生成模型在研究正常和病理条件下红系分化的机制方面非常有用。在这里，我们描述了一种从脐血来源的造血干细胞（HSCs）开始的红系液体培养系统。在红系分化条件下培养 HSCs 超过 50 天，在最佳条件下可在 50 天内实现超过 10 倍的扩增。通过细胞形态、流式细胞术和造血集落测定对均一性红细胞进行了特征描述。此外，通过与人胎肝基质细胞共培养可改善终末红系成熟。共培养的红细胞经历了多次成熟事件，包括体积减小、糖蛋白 A 表达增加和核浓缩。这一过程导致多达 80%的细胞中出现固缩核的挤出。重要的是，它们在进一步成熟后具有表达成人定型β-珠蛋白链的能力。我们还表明，脐血分化的红细胞（RBCs）的氧平衡曲线与正常 RBCs 相当。大量且纯度高的脐血来源的红细胞可用于红系发育的基础研究，并为未来生产可用于输血的可用 RBCs 提供了基础。
