Non-transmissible measles virus vector with segmented RNA genome establishes different types of iPSCs from hematopoietic cells. Non-transmissible measles virus vector with segmented RNA genome establishes different types of iPSCs from hematopoietic cells

We established iPSCs from CD34+ HPCs or CD3+ T cells by transducing measles virus vectors encording reprogramming genes. HPC-derived iPSCs and T-cell-derived iPSCs showed naive-like and primed pluripotent cell properties, respectively. Overall design: CD34+ and CD3+ cells were sorted from cord blood and transduced with measles virus vectors encoding OCT3/4, SOX2, KLF4 and L-MYC. The cells were cultured in regular human ESC culture media for 2-3 weeks. After development of colonies, the cells were harvested and dissociated, then cultured with in naive media or human ESC culture media. After several passages, total cell RNA was harvested and subjected to RNA seq.

本研究通过转导编码重编程基因的麻疹病毒载体，从CD34阳性造血祖细胞（CD34+ hematopoietic progenitor cells, HPCs）或CD3阳性T细胞中构建诱导多能干细胞（induced pluripotent stem cells, iPSCs）。造血祖细胞来源的诱导多能干细胞与T细胞来源的诱导多能干细胞，分别呈现类幼稚态与始发态多能细胞的表型特性。整体实验设计：从脐带血中分选获得CD34阳性与CD3阳性细胞，采用携带OCT3/4、SOX2、KLF4及L-MYC基因的麻疹病毒载体进行转导；将细胞置于常规人类胚胎干细胞（embryonic stem cells, ESC）培养基中培养2~3周。待细胞克隆形成后，收集细胞并进行解离，随后分别接种于幼稚态培养基或常规人类胚胎干细胞培养基中继续培养。经过数次传代后，提取细胞总RNA并开展RNA测序（RNA sequencing, RNA-seq）。