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首页> 外文期刊>BioMed research international >In Vitro Large Scale Production of Human Mature Red Blood Cells from Hematopoietic Stem Cells by Coculturing with Human Fetal Liver Stromal Cells
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In Vitro Large Scale Production of Human Mature Red Blood Cells from Hematopoietic Stem Cells by Coculturing with Human Fetal Liver Stromal Cells

机译：通过用人胎儿肝脏基质细胞与造血干细胞从造血干细胞进行体外大规模生产人类成熟红细胞

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摘要

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 109-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 beta-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.
机译：人类红细胞的体外模型可用于研究正常和病理条件下红细胞分化的机制。在这里，我们描述了从脐带血源性造血干细胞（HSC）开始的红细胞液体培养系统。在红细胞分化条件下培养HSC超过50天，在最佳条件下在50天内产生超过109倍的膨胀。通过细胞形态，流式细胞术和造血菌落测定表征均匀红细胞细胞。此外，通过用人胎儿肝脏基质细胞缓解末端红细胞成熟。共禾红外细胞接受多个成熟事件，包括尺寸的减小，糖蛋白的表达增加，核凝结。该过程导致挤出Pycnotic核在高达80％的细胞中。重要的是，它们在进一步成熟时具有表达成人最终β-珠蛋白链的能力。我们还表明，脐带血分症红细胞（RBC）的氧平衡曲线与正常RBC相当。从脐带血中产生的红细胞和RBC的大量和纯度使得该方法可用于红细胞发展的基本研究，并且还为未来生产可用RBC的输血提供了基础。

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