Efficient in vitro labeling of human neural precursor cells with superparamagnetic iron oxide particles: Relevance for in vivo cell tracking
Efficient in vitro labeling of human neural precursor cells with superparamagnetic iron oxide particles: Relevance for in vivo cell tracking
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DOI：
10.1634/stemcells.2007-0251
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发表时间：
2008-02-01
期刊：
影响因子：
5.2
通讯作者：
Gritti, Angela
中科院分区：
文献类型：
--
作者：
Neri, Margherita;Maderna, Claudio;Gritti, Angela
最近的研究提出了通过移植体外扩增的神经前体细胞（NPC）和/或其后代来替代受损或丢失的神经细胞的诱人可能性。超顺磁性氧化铁 (SPIO) 标记细胞的磁共振 (MR) 追踪是一种在活体动物纵向研究中追踪移植细胞的非侵入性技术。 SPIO 可以有效标记小鼠 NPC 和人类间充质或造血干细胞。然而，基于 SPIO 的标记人类神经前体细胞 (hNPC) 方案的验证尚未得到广泛解决。在这里，我们报告了使用两种 SPIO（Sinerem 和 Endorem）来标记在体外表现出真正的干细胞特征的人类 hNPC 的优化方案的开发和验证。需要对两种 SPIO 进行仔细滴定，以设定产生有效细胞标记而不损害细胞存活、增殖、自我更新和多能性的条件。体内磁共振成像（MRI）结合组织学和共聚焦显微镜表明，在成年小鼠大脑移植后的短期内可以有效地检测到少量（5 x 10（3）至1 x 10（4））的活的SPIO标记的hNPC，并且可以在纵向研究中跟踪至少1个月。通过使用这种方法，我们还阐明了供体细胞死亡对 MR 信号的影响。本研究描述了一种简单的方案，使用优化的低剂量 SPIO 来标记人类来源的 NPC，并证明了在大脑移植后对标记细胞进行无创成像的可行性；它还证明了必须考虑的技术的潜在局限性，特别是从基于神经细胞的临床应用的角度来看。
Recent studies have raised appealing possibilities of replacing damaged or lost neural cells by transplanting in vitro-expanded neural precursor cells (NPCs) and/or their progeny. Magnetic resonance (MR) tracking of superparamagnetic iron oxide (SPIO)-labeled cells is a non-invasive technique to track transplanted cells in longitudinal studies on living animals. Murine NPCs and human mesenchymal or hematopoietic stem cells can be efficiently labeled by SPIOs. However, the validation of SPIO-based protocols to label human neural precursor cells (hNPCs) has not been extensively addressed. Here, we report the development and validation of optimized protocols using two SPIOs (Sinerem and Endorem) to label human hNPCs that display bona ride stem cell features in vitro. A careful titration of both SPIOs was required to set the conditions resulting in efficient cell labeling without impairment of cell survival, proliferation, self-renewal, and multipotency. In vivo magnetic resonance imaging (MRI) combined with histology and confocal microscopy indicated that low numbers (5 x 10(3) to 1 x 10(4)) of viable SPIO-labeled hNPCs could be efficiently detected in the short term after transplantation in the adult murine brain and could be tracked for at least 1 month in longitudinal studies. By using this approach, we also clarfied the impact of donor cell death to the MR signal. This study describes a simple protocol to label NPCs of human origin using SPIOs at optimized low dosages and demonstrates the feasibility of noninvasive imaging of labeled cells after transplantation in the brain; it also evidentiates potential limitations of the technique that have to be considered, particularly in the perspective of neural cell-based clinical applications.