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.
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.
Synsnerven giver et ideelt sted for centralnervesystemet (CNS) regenerativ forskning, herunder oftalmologiske forhold som optisk neuritis, grøn stær og traumer. Injektioner af en række stamceller har enten vist virkning eller vist lovende i at erstatte tabt myelin, øge axonal tælle og / eller forebyggelse af degenerative sygdomme. 1,2
Den menneskelige synsnerven indeholder ca. 1,2 mio parallelle axoner rejser fra nethinden til chiasm med en diameter på ca. 3,0-3,5 mm. 3 For at modellere humane sygdomme i laboratoriet, er rotten blevet brugt hyppigt. Den voksne rotte synsnerve indeholder ca. 100.000 axoner inden for en diameter på ca. 0,5 mm. 4 En af de største begrænsninger i CNS regenerativ forskning er direkte udbenet adgang. Komplikationer og kirurgiske risici for dyret er højere, når kraniet eller ryghvirvler fjernes. Svarende til fordelene vedminimalt invasive metoder i rygsøjlen, 5 direkte optiske nerve injektioner uden at åbne kraniet tilbyder reduceret komplikationer og en hurtigere genopretning.
Denne teknik har været anvendt i tidligere undersøgelser. 6 I dette håndskrift og ledsagende video viser vi en minimalt invasiv procedure til at injicere stamceller i rotter synsnerve.
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…
The authors have nothing to disclose.
This study was supported by NeuralStem, Inc., and Johns Hopkins Project RESTORE.
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 |