微创技术注入到老鼠视神经
Summary
Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.
Abstract
The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.
Introduction
视神经为中枢神经系统（CNS）再生的研究，包括眼科疾病如视神经炎，青光眼和创伤的理想地点。的各种干细胞注射要么证明疗效或显示出希望在替换丢失髓磷脂，增加轴突计数和/或预防变性疾病。1,2-
人类视神经含有约120万平行轴突从视网膜到一个直径约为3.0-3.5毫米的视交叉行进。3为了模拟人类疾病在实验室中，老鼠被频繁使用。成年大鼠视神经包含大约100,000轴突直径大约0.5mm范围内。4之一在中枢神经系统的主要限制再生的研究是直接去骨访问。并发症和手术风险的动物都在颅骨或脊椎被删除更高。类似的好处微创方法在脊柱，5个直属视神经注射，无需打开颅骨报价减少并发症和更快的恢复。
该技术已在以前的研究。6在这个手稿和影随行，我们展示了微创方法，以干细胞注入到老鼠视神经。
Protocol
Representative Results
Discussion
Direct injection into the optic nerve of stem cells or other products intended to facilitate regeneration provides a convenient model compared to other means of injections into the CNS. This technique takes less time, requires less total anesthesia, avoids drilling or removing skull or bone tissue, reduces complications rates and allows for more rapid recovery following surgery.
The most critical steps in this protocol include: 1. Adequate hemostasis in the surgical field to allow clear visua…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This study was supported by NeuralStem, Inc., and Johns Hopkins Project RESTORE.
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Lewis rat | Charles River | 4 | Any rat strain will work. |
| Anesthesia machine | Surgivet | CDS9000 | CDS 9000 Small Animal Anesthesia Machine – Pole Mount |
| Infusion pump | Stoelting | 53129 | |
| Dissection microscope | National Optical | 409-411-1105 | |
| Fiber-optic light source | Fisher Scientific | 12-562-21 | |
| Dissection and Stereotaxic Instrument | Stoelting | 51400 | |
| Pipette Puller | Kopf | 750 | |
| Pipettes | World Precision Instruments | 18150-6 | |
| Disposable scalpel blades | Harvard Apparatus | 810-15-021 | |
| Iridectomy scissors | Electron Microscopy Sciences | Uniband LA-4XF |
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