Maintenance of proliferative capacity and retroviral transduction efficiency of human fetal CD38(-)/CD34(++) stem cells.
Maintenance of proliferative capacity and retroviral transduction efficiency of human fetal CD38(-)/CD34(++) stem cells.
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维持人胎儿CD38(-)/CD34( )干细胞的增殖能力和逆转录病毒转导效率。
DOI：
10.1089/scd.2006.15.97
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发表时间：
2006
影响因子：
4
通讯作者：
Bárcena,Alicia
中科院分区：
文献类型：
--
作者：
Muench,MarcusO;Ohkubo,Tatsuo;Smith,ClaytonA;Suskind,DavidL;Bárcena,Alicia
用于胎儿造血干细胞的有效转导和扩增的方法可以导致用于血细胞病症和酶缺乏的新颖的子宫内疗法。在这里，我们描述了一种新的检测方法，以快速测量细胞因子对从人妊娠中期肝脏分离的CD 38-CD 34 ++谱系细胞中发现的原始祖细胞和干细胞的分化或扩增的影响。重要的是，支持克隆祖细胞扩增的条件降低了它们的增殖能力。巨核细胞生长和发育因子和粒细胞-巨噬细胞集落刺激因子的组合保持增殖潜力，同时也产生中等水平的祖细胞扩增。逆转录病毒转导使用基于莫洛尼鼠白血病病毒的载体实现。新鲜分离的候选干细胞可以通过暴露于病毒1小时并离心以辅助转导而以几乎17%的效率转导。在培养1天后，这增加到平均35.5%的转导效率。此外，从胎儿胎盘血中分离的候选干细胞的转导效率为33.0%。这些发现鼓励进一步研究子宫外基因治疗的可行性，即胎儿细胞从循环中分离，转导，并在子宫外扩增，然后再返回胎儿。
Methods for the efficient transduction and expansion of fetal hematopoietic stem cells could lead to novel in utero therapies for blood cell disorders and enzymatic deficiencies. Here we describe a new assay to measure rapidly the effects of cytokines on the differentiation or expansion of primitive progenitors and stem cells found among CD38–CD34++lineage–cells isolated from human midgestation liver. Importantly, conditions that otherwise supported the expansion of clonogenic progenitors reduced their proliferative capacity. A combination of megakaryocyte growth and development factor and granulocyte-macrophage colony-stimulating factor maintained proliferative potential while also yielding an intermediate level of progenitor expansion. Retroviral transduction was achieved using Moloney murine leukemia virus-based vectors. Freshly isolated candidate stem cells could be transduced at almost 17% efficiency by a 1-h exposure to virus with centrifugation to aid transduction. This was increased to a mean 35.5% transduction efficiency after 1 day of culture. Additionally, the transduction efficiency of candidate stem cells isolated from fetal placental blood was 33.0%. These findings encourage further investigation into the feasibility of ex utero gene therapy whereby fetal cells are isolated from the circulation, transduced, and expanded ex utero before being returned to the fetus.