Molecular and Magnetic Resonance Imaging of Human Embryonic Stem Cell-Derived Neural Stem Cell Grafts in Ischemic Rat Brain
Molecular and Magnetic Resonance Imaging of Human Embryonic Stem Cell-Derived Neural Stem Cell Grafts in Ischemic Rat Brain
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DOI：
10.1038/mt.2009.104
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发表时间：
2009-07-01
影响因子：
12.4
通讯作者：
Steinberg, Gary K.
中科院分区：
文献类型：
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作者：
Daadi, Marcel M.;Li, Zongjin;Steinberg, Gary K.
移植干细胞的实时成像对于了解它们在体内与宿主环境的相互作用、追踪细胞命运和功能以及在临床环境中成功递送和安全监测至关重要。在这项研究中，我们使用生物发光（BLI）和磁共振成像（MRI）来观察移植的人胚胎干细胞（hESC）来源的人神经干细胞（hNSCs）在中风损伤大鼠脑中的命运。用携带双融合（DF）报告基因的慢病毒载体对hNSCs进行基因工程改造，该报告基因稳定表达增强型绿色荧光蛋白（eGFP）和萤火虫荧光素酶（fLuc）报告基因。hNSCs是自我再生的，多能的，并表达神经干细胞的标记物。移植后2个月，通过MRI和BLI无创追踪细胞存活，并进行组织学证实。移植的GFP（+）细胞的电生理记录和免疫电镜显示了连通性。移植的hNSCs分化为神经元、发生髓鞘再生的脑卒中区域的少突胶质细胞和向脑卒中受损血管延伸的星形胶质细胞。我们的数据表明，BLI和MRI模式的结合提供了可靠的细胞命运实时监测。
Real-time imaging of transplanted stem cells is essential for understanding their interactions in vivo with host environments, for tracking cell fate and function and for successful delivery and safety monitoring in the clinical setting. In this study, we used bioluminescence (BLI) and magnetic resonance imaging (MRI) to visualize the fate of grafted human embryonic stem cell (hESC)-derived human neural stem cells (hNSCs) in stroke-damaged rat brain. The hNSCs were genetically engineered with a lentiviral vector carrying a double fusion (DF) reporter gene that stably expressed enhanced green fluorescence protein (eGFP) and firefly luciferase (fLuc) reporter genes. The hNSCs were self-renewable, multipotent, and expressed markers for neural stem cells. Cell survival was tracked noninvasively by MRI and BLI for 2 months after transplantation and confirmed histologically. Electrophysiological recording from grafted GFP(+) cells and immuno-electronmicroscopy demonstrated connectivity. Grafted hNSCs differentiated into neurons, into oligodendrocytes in stroke regions undergoing remyelination and into astrocytes extending processes toward stroke-damaged vasculatures. Our data suggest that the combination of BLI and MRI modalities provides reliable real-time monitoring of cell fate.