High Transduction Efficiency of Human Amniotic Fluid Stem Cells Mediated by Adenovirus Vectors
High Transduction Efficiency of Human Amniotic Fluid Stem Cells Mediated by Adenovirus Vectors
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DOI：
10.1089/scd.2007.0188
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发表时间：
2008-10-01
影响因子：
4
通讯作者：
Tomanin, Rosella
中科院分区：
文献类型：
--
作者：
Grisafi, Davide;Piccoli, Martina;Tomanin, Rosella
在过去的几年里，一些研究表明，从羊水中培养出具有各种潜能的祖细胞是可能的。羊膜穿刺术是一种被广泛接受的产前诊断方法；它对母亲和胎儿的风险都很低，并克服了通常与其他来源相关的伦理问题。最近，我们描述了羊水干细胞(AFS)，因为他们的能力，分化为不同的谱系，可能是一个很好的候选治疗应用。为了达到基因治疗的目的，将治疗性基因导入人骨髓间充质干细胞，或直接注射入靶组织。本研究的目的是探索腺病毒载体转导人骨髓间充质干细胞的可行性，并确定转导后的干细胞是否具有向不同系分化的能力。在此，我们证明了第一代腺病毒载体可以有效地感染HAFS细胞。此外，我们还证明了两个不同的标记基因LacZ和EGFP的感染和表达对细胞的表型和分化潜能没有影响。特别是，在未分化状态下，Hafs细胞继续表达转基因和干细胞标记OCT4和SSEA4。当感染细胞在间质条件下培养时，仍可分化为表达谱系特异性基因的骨细胞和脂肪细胞。这些初步发现表明，腺病毒可能有助于工程多能干细胞群体，并可能用于广泛的基因治疗。
In the last few years some studies have shown the possibility of deriving progenitors with various potential from the amniotic fluid. Amniocentesis is a widely accepted method for prenatal diagnosis; it is associated with low risk both for the mother and the fetus and overcomes the ethical problems commonly associated to other sources. Recently we have described that amniotic fluid stem (AFS) cells, for their ability to differentiate to various lineages, could represent a good candidate for therapeutic applications. For gene therapy purposes human AFS (hAFS) cells should be genetically modified with a therapeutic gene and delivered systematically or injected directly into the tissue of interest.The aim of this study was to investigate the feasibility of transducing hAFS cells with adenoviral vectors and to determine whether transduced stem cells retain the ability to differentiate into different lineages. Herein, we showed that hAFS cells could be efficiently infected by first generation adenovirus vectors. In addition, we demonstrated that infection and expression of two different marker genes, LacZ and EGFP, have no effect on cells phenotype and differentiation potential. In particular, on undifferentiated status, hAFS cells continued to express both the transgenes and stemness cell markers OCT4 and SSEA4. When cultured under mesenchymal conditions, infected cells could still differentiate into osteocytes and adipocytes expressing lineage specific genes.These preliminary findings suggest that adenovirus may be useful to engineer populations of pluripotent stem cells, which may be used in a wide range of gene therapy treatments.