Generation of human induced pluripotent stem cells from cord blood cells.
Generation of human induced pluripotent stem cells from cord blood cells.
复制标题
DOI：
--
复制
发表时间：
2011
影响因子：
--
通讯作者：
N. Nishishita;C. Takenaka;N. Fusaki;S. Kawamata
中科院分区：
文献类型：
--
作者：
N. Nishishita;C. Takenaka;N. Fusaki;S. Kawamata
我们报道了用携带重编程因子Oct3/4、SOX2、KLF4和c-myc的仙台病毒(SeV)载体从新鲜的人脐带血(CB)细胞中安全有效地产生iPS细胞。SeV载体是一种没有DNA时相的单链RNA病毒，选择性地感染与造血祖细胞相对应的新鲜分离的CD34+CD45low+部分CB细胞。在SeV感染后18天左右，从新鲜分离的1×104 CD34+CB细胞中产生了大约20个ES细胞样集落，并被选择用于传代，以减少剩余SeV感染细胞的频率。用抗血凝素神经氨酸酶(HN)的单抗进行免疫染色和RT-PCR分析，证实病毒载体经过多次传代后被完全消除。选择5个ES细胞样克隆，检测其体外三胚层分化潜能和畸胎瘤形成能力。从CB细胞培养出非整合的仙台病毒(SeV)iPS细胞可能是未来提供异基因iPS细胞来源治疗的重要一步。
We report that iPS cells can be safely and effectively generated from fresh human cord blood (CB) cells with Sendai virus (SeV) vector carrying reprogramming factors OCT3/4, SOX2, KLF4, and c-MYC. The SeV vector is a single strand RNA virus having no DNA phase, and selectively infects the freshly isolated CD34+ CD45low+ fraction of CB cells corresponding to hematopoietic progenitors. Approximately twenty ES cell-like colonies emerged from 1 x 104 freshly isolated CD34+ CB cells around 18 days after SeV infection and were selected for passage to reduce the frequency of the remaining SeV-infected cells. The complete elimination of viral constructs was confirmed after several passages by immunostaining with monoclonal antibody against hemagglutinin-neuraminidase (HN) and by RT-PCR analysis. Five ES cell-like clones were selected to examine their in vitro potential for three germ layer differentiation and their capacity for teratoma formation. Generation of non-integrating Sendai virus (SeV) iPS cells from CB cells may be an important step to provide allogeneic iPS cell-derived therapy in the future.