Department of Medicine and Institute for Human Genetics, University of California, San Francisco, San Francisco, CA 94143, USA.
Stem Cells Transl Med. 2013 Aug;2(8):558-66. doi: 10.5966/sctm.2013-0006. Epub 2013 Jul 11.
The discovery of induced pluripotent stem cells (iPSCs) holds great promise for regenerative medicine since it is possible to produce patient-specific pluripotent stem cells from affected individuals for potential autologous treatment. Using nonintegrating cytoplasmic Sendai viral vectors, we generated iPSCs efficiently from adult mobilized CD34⁺ and peripheral blood mononuclear cells. After 5-8 passages, the Sendai viral genome could not be detected by real-time quantitative reverse transcription-polymerase chain reaction. Using the spin embryoid body method, we showed that these blood cell-derived iPSCs could efficiently be differentiated into hematopoietic stem and progenitor cells without the need of coculture with either mouse or human stromal cells. We obtained up to 40% CD34⁺ of which ~25% were CD34⁺/CD43⁺ hematopoietic precursors that could readily be differentiated into mature blood cells. Our study demonstrated a reproducible protocol for reprogramming blood cells into transgene-free iPSCs by the Sendai viral vector method. Maintenance of the genomic integrity of iPSCs without integration of exogenous DNA should allow the development of therapeutic-grade stem cells for regenerative medicine.
诱导多能干细胞（iPSCs）的发现为再生医学带来了巨大的希望，因为有可能从受影响的个体中产生患者特异性的多能干细胞，用于潜在的自体治疗。我们使用非整合的细胞质仙台病毒载体，从成年动员的 CD34+和外周血单个核细胞中高效地产生了 iPSCs。经过 5-8 个传代，实时定量逆转录聚合酶链反应检测不到仙台病毒基因组。使用旋转胚胎体方法，我们表明这些血细胞衍生的 iPSCs 可以在不需要与小鼠或人基质细胞共培养的情况下有效地分化为造血干细胞和祖细胞。我们获得了高达 40%的 CD34+细胞，其中约 25%为 CD34+/CD43+造血前体细胞，可轻易分化为成熟血细胞。我们的研究通过仙台病毒载体方法证明了一种可重复的方案，可将血细胞重编程为无转基因的 iPSCs。维持 iPSCs 的基因组完整性而不整合外源 DNA，应允许开发用于再生医学的治疗级别的干细胞。
