Sıçan Optik Sinir içine Enjeksiyon için minimal invaziv Tekniği
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
Optik sinir merkezi sinir sistemi (MSS) gibi optik nevrit, glokom ve travma olarak göz koşullar da dahil olmak üzere rejeneratif araştırma için ideal bir konum sağlar. Kök hücrelerin çeşitli enjeksiyonları ya aksonal sayısı artan ve / veya dejeneratif hastalıkların önlenmesi, etkinlik göstermiştir veya kayıp miyelin değiştirilmesi söz göstermiştir. 1,2
insan optik sinir yaklaşık 3.0-3.5 mm'lik bir çapa sahip kiazmada retinadan seyahat eden yaklaşık 1.200.000, paralel aksonlar içerir. 3 Laboratuarda insan hastalıklarının modellemek için, sıçan sık kullanılmaktadır. yetişkin sıçan optik sinir yaklaşık 0,5 mm çapında içinde yaklaşık 100.000 aksonlar içerir. MSS önemli sınırlamalar 4 biri rejeneratif araştırma direkt kemiksiz erişimi olduğunu. Hayvan komplikasyonlar ve cerrahi riskler kafatası veya omurlar kaldırıldığında yüksektir. Faydaları Benzeromurgada minimal invazif yaklaşımlar, kafatası teklif azaltılmış komplikasyonları ve daha hızlı bir toparlanma açmadan 5 direk optik sinir enjeksiyonları.
Bu teknik daha önceki çalışmalarda kullanılmıştır. 6 Bu yazıda ve beraberindeki video, sıçan optik sinir içine kök hücreler enjekte etmek için minimal invaziv prosedür göstermek.
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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