In vivo imaging of engrafted neural stem cells: its application in evaluating the optimal timing of transplantation for spinal cord injury
In vivo imaging of engrafted neural stem cells: its application in evaluating the optimal timing of transplantation for spinal cord injury
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DOI：
10.1096/fj.05-4082fje
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发表时间：
2005-09-01
期刊：
影响因子：
4.8
通讯作者：
Okano, H
中科院分区：
文献类型：
--
作者：
Okada, S;Ishii, K;Okano, H
神经干/祖细胞（NSPCs）有望在脊髓损伤后的神经组织替代治疗中发挥作用。然而，了解移植的NSPCs的存活时间并确定从移植部位迁移的程度对于优化治疗方案至关重要。在这里，我们使用体内生物发光成像来非侵入性地评估移植的NSPCs在活体动物损伤部位的存活和停留时间，并且我们使用组织学分析来评估细胞整合和形态。第三代慢病毒载体使荧光素酶和绿色荧光蛋白的变体在原代培养的NSPCs中能够有效转导和稳定表达。这些细胞的信号在移植到C57 BL/6 J小鼠的损伤脊髓后长达10个月或更长时间内都是可检测的。组织学和功能数据支持成像数据，并表明NSPC移植的时间可能是整合细胞的命运和功能的关键决定因素，因为细胞存活和迁移取决于相对于损伤的移植时间。通过成像可以大大加速和改进细胞疗法的优化，并且本研究中的方法可以广泛应用于再生医学的各个研究领域，包括移植研究。
Neural stem/progenitor cells (NSPCs) hold promise in neural tissue replacement therapy after spinal cord injury. However, understanding the survival time of grafted NSPCs and determining the extent of migration away from transplantation sites are essential for optimizing treatment regimens. Here, we used in vivo bioluminescence imaging to noninvasively assess the survival and residence time of transplanted NSPCs at the injury sites in living animals, and we used histologic analyses to assess cell integration and morphology. Third-generation lentiviral vectors enabled efficient transduction and stable expression of both luciferase and a variant of green fluorescent protein in primary cultured NSPCs. Signals from these cells were detectable for up to 10 months or more after transplantation into the injured spinal cords of C57BL/6J mice. Histological and functional data supported the imaging data and suggest that the timing of NSPC transplantation may be a key determinant of the fates and function of integrated cells since cell survival and migration depended on the time of transplantation relative to injury. Optimization of cell therapies can be greatly accelerated and refined by imaging, and the methods in the present study can be widely applied to various research fields of regeneration medicine, including transplantation study.