In vivo bioluminescent imaging of virus-mediated gene transfer and transduced cell transplantation in the intervertebral disc
In vivo bioluminescent imaging of virus-mediated gene transfer and transduced cell transplantation in the intervertebral disc
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DOI：
10.1097/00007632-200404150-00004
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发表时间：
2004-04-15
期刊：
影响因子：
3
通讯作者：
Anderson, DG
中科院分区：
文献类型：
--
作者：
Leo, BM;Li, XD;Anderson, DG
研究设计.本文介绍的工作使用小动物模型来证明体内生物发光成像研究退行性椎间盘疾病的可行性和有用性。确定体内生物发光成像监测啮齿动物模型椎间盘内遗传修饰细胞的时空表达的实用性。脊柱中基因工程细胞的非侵入性成像具有允许在不杀死动物的情况下跟踪事件的优点，并且可以用于跟踪特定治疗的时间过程。结果上使用的Sprague-Dawley大鼠在实验研究中，荧光素酶报告基因被传递到腰椎间盘通过腺病毒介导的或基于细胞的转移技术，以证明随着时间的推移，非侵入性地监测基因表达的可行性。组织培养，椎间盘手术，并在体内生物发光成像。用含有荧光素酶报告基因的腺病毒原位注射大鼠的椎间盘，或植入离体转导并包含在生物可吸收载体中的脂肪、骨髓或椎间盘细胞。在几个时间点用体内生物发光成像评估结果。通过长达14天的成像获得了11只动物的数据。据我们所知，这是第一次描述体内生物发光成像研究脊柱条件。我们已经表征了通过离体转染用Ad-luc病毒转导的三种细胞类型的相对表达，然后在大鼠脊柱中进行细胞植入，并将它们彼此进行比较，并与Ad-luc腺病毒原位直接感染进行比较。我们的研究结果证明了在脊柱中追踪基因改变细胞的可行性。这种技术有可能被用于非侵入性地跟踪在椎间盘研究中使用的小动物的脊柱内的治疗基因的命运和表达。
Study Design. Work presented here used a small animal model to demonstrate the feasibility and usefulness of in vivo bioluminescent imaging to studying degenerative disc disease.Objectives. To determine the utility of in vivo bioluminescent imaging to monitor the temporal and spatial expression of genetically modified cells within the intervertebral disc of a rodent model.Summary of the Background Data. Noninvasive imaging of genetically engineered cells in the spine has the advantage of allowing events to be tracked without killing the animal and can be used to follow the time course of a particular therapy. Results are presented on the use of Sprague-Dawley rats in experimental studies in which the luciferase reporter gene was delivered to the lumbar intervertebral disc through adenovirus-mediated or cell-based transfer techniques to demonstrate the feasibility to monitor gene expression noninvasively over time.Methods. Tissue culture, disc surgery, and in vivo bioluminescent imaging were used. The intervertebral disc of the rat was either injected in situ with an adenovirus containing the luciferase reporter gene or implanted with fat, bone marrow or intervertebral disc cells transduced ex vivo and contained in a bioresorbable carrier. Results were assessed with in vivo bioluminescent imaging at several time points.Conclusion. Data from 11 animals were obtained with imaging up to 14 days. To our knowledge, this is the first description of in vivo bioluminescence imaging to study spinal conditions. We have characterized the relative expression of three cell types transduced with the Ad-luc virus by ex vivo transfection followed by cell implantation in the rat spine and compared them to one another and to direct infection of Ad-luc adenovirus in situ. Our results demonstrate the feasibility of tracing genetically altered cells in the spine. This technique has the potential to be used to noninvasively track the fate and expression of therapeutic genes within the spine of small animals used in disc research.