Center for Regenerative Medicine in Barcelona, Dr. Aiguader, Barcelona, Spain.
Nat Protoc. 2010 Apr;5(4):811-20. doi: 10.1038/nprot.2010.16. Epub 2010 Apr 1.
Induced pluripotent stem cells (iPSC) provide an invaluable resource for regenerative medicine as they allow the generation of patient-specific progenitors with potential value for cell therapy. However, in many instances, an off-the-shelf approach is desirable, such as for cell therapy of acute conditions or when the patient's somatic cells are altered as a consequence of a chronic disease or aging. Cord blood (CB) stem cells appear ideally suited for this purpose as they are young cells expected to carry minimal somatic mutations and possess the immunological immaturity of newborn cells; additionally, several hundred thousand immunotyped CB units are readily available through a worldwide network of CB banks. Here we present a detailed protocol for the derivation of CB stem cells and how they can be reprogrammed to pluripotency by retroviral transduction with only two factors (OCT4 and SOX2) in 2 weeks and without the need for additional chemical compounds.
诱导多能干细胞(iPSC)为再生医学提供了宝贵的资源,因为它们可以生成具有细胞治疗潜力的患者特异性祖细胞。然而,在许多情况下,需要采用现成的方法,例如治疗急性病症,或者当患者的体细胞由于慢性疾病或衰老而发生改变时。脐带血(CB)干细胞似乎非常适合这种用途,因为它们是年轻的细胞,预期携带最小的体细胞突变,并具有新生儿细胞的免疫不成熟性;此外,通过全球脐带血库网络,可轻易获得几十万份免疫定型的 CB 单位。在这里,我们提供了一个详细的 CB 干细胞的衍生方案,以及如何通过逆转录病毒转导仅用两周时间和两种因子(OCT4 和 SOX2)将其重编程为多能性,而无需添加其他化合物。