Lu L, Li Z H, He J, Ge Y, Rice S, Broxmeyer H E
Department of Microbiology, Walther Oncology Center, Indiana University School of Medicine, Indianapolis 46202-5121, USA.
J Leukoc Biol. 1998 Mar;63(3):389-94. doi: 10.1002/jlb.63.3.389.
In this study, we tested the capacity to change the differentiation profile of progenitor cells by retroviral-mediated transduction of EpoR cDNA into one of the paired daughter cells derived from single CD34(3+) CB cells. Our results show that for the non-viral-treated daughter cells, the majority (99.6%) formed the same colony type. However, with cells transduced with viral vectors, 7.1% of the daughter cells transduced with the EpoR cDNA formed either a burst forming unit-erythroid (BFU-E) or a colony-forming unit-granulocyte, macrophage, erythroid, megakaryocyte (CFU-GEMM) colony compared to the other daughter cell transduced with viral supernatant lacking EpoR cDNA, which formed either a colony-forming unit granulocyte-macrophage (CFU-GM) or a high proliferative potential-colony forming cell (HPP-CFC) colony. Expression of the transduced EpoR cDNA was confirmed in individual colonies by RT-PCR analysis. These results substantiate in a more rigorous fashion our previous results that it is possible to change the Epo-responsive differentiation profile of progenitor cells by transduction into these cells of an EpoR cDNA and this change was apparent only in daughter cells derived from single CD34(3+) kit+ cells transduced with EpoR cDNA.
在本研究中，我们测试了通过逆转录病毒介导将促红细胞生成素受体（EpoR）cDNA转导至源自单个CD34(3+)脐血（CB）细胞的一对子代细胞之一中，从而改变祖细胞分化谱的能力。我们的结果表明，对于未进行病毒处理的子代细胞，大多数（99.6%）形成相同的集落类型。然而，在用病毒载体转导的细胞中，与用缺乏EpoR cDNA的病毒上清液转导的另一个子代细胞相比，转导了EpoR cDNA的子代细胞中有7.1%形成了红系爆式集落形成单位（BFU-E）或粒-巨噬-红-巨核系集落形成单位（CFU-GEMM）集落，后者形成了粒-巨噬系集落形成单位（CFU-GM）或高增殖潜能集落形成细胞（HPP-CFC）集落。通过逆转录-聚合酶链反应（RT-PCR）分析在单个集落中证实了转导的EpoR cDNA的表达。这些结果以更严格的方式证实了我们之前的结果，即通过将EpoR cDNA转导至这些细胞中有可能改变祖细胞对促红细胞生成素的反应性分化谱，并且这种变化仅在转导了EpoR cDNA的单个CD又(3+)试剂盒+细胞衍生的子代细胞中明显。
