Lu Shi-Jiang, Feng Qiang, Park Jennifer S, Lanza Robert
Advanced Cell Technology, Worcester, MA, USA.
Methods Mol Biol. 2010;636:105-21. doi: 10.1007/978-1-60761-691-7_7.
Human embryonic stem cells (hESC) represent a new source of stem cells that can be propagated and expanded in vitro indefinitely, providing a potentially inexhaustible and donorless source of cells for human therapy. The ability to create banks of hESC lines with matched or reduced incompatibility could potentially reduce or eliminate the need for immunosuppressive drugs and/or immunomodulatory protocols altogether, for example, O-type RhD(-) lines for generation of universal red blood cells (RBC). Hematopoietic differentiation of hESCs has been extensively investigated in vitro, and hematopoietic precursors as well as differentiated progeny representing erythroid, myeloid, macrophage, megakaryocytic, and lymphoid lineages have been identified in differentiating hESC cultures. Previous studies also generated primitive erythroid cells from hESCs by embryoid body (EB) formation and coculturing with stromal cells. However, the efficient and controlled differentiation of hESCs into homogeneous RBC populations with oxygen-carrying capacity has not been previously achieved. In this chapter, we describe a robust system that can efficiently generate large numbers of hemangioblasts from multiple hESC lines using well-defined conditions and produce functional homogeneous RBCs with oxygen-carrying capacity in large scale. The homogeneous erythroid cells can be used for further mechanism studies.
人类胚胎干细胞（hESC）代表了一种新的干细胞来源，它可以在体外无限期地增殖和扩增，为人类治疗提供了一个潜在的取之不尽且无需供体的细胞来源。创建具有匹配或降低不相容性的hESC系库的能力可能会减少或完全消除对免疫抑制药物和/或免疫调节方案的需求，例如，用于生成通用红细胞（RBC）的O型RhD（-）系。hESC的造血分化已在体外进行了广泛研究，并且在分化的hESC培养物中已鉴定出造血前体以及代表红系、髓系、巨噬细胞、巨核细胞和淋巴系的分化后代。先前的研究还通过胚状体（EB）形成和与基质细胞共培养从hESC中产生了原始红细胞。然而，此前尚未实现将hESC高效且可控地分化为具有携氧能力的均匀RBC群体。在本章中，我们描述了一种强大的系统，该系统可以使用明确的条件从多个hESC系高效地产生大量成血管细胞，并大规模生产具有携氧能力的功能性均匀RBC。这些均匀的红系细胞可用于进一步的机制研究。
