Directed retinal nerve cell growth for use in a retinal prosthesis interface.
OBJECTIVE
Retinal prosthetic devices that use microelectrode arrays to stimulate retinal nerve cells may provide a viable treatment for degenerative retinal diseases. Current devices are based on electrical field-effect stimulation of remaining functional neural elements. However, the distance between target neurons and electrodes limits the potential density of electrodes and the ability to stimulate specific types of retinal neurons that contribute to visual perceptions. This study was conducted to investigate the use of microcontact printing (muCP) to direct cultured or explant retinal ganglion cell (RGC) neurites to precise and close stimulation positions and to evaluate the cell types that grow from a retinal explant.


METHODS
RGCs and whole retinal explants were isolated from postnatal day-7 Sprague-Dawley rats using immunopanning purification and microdissection, respectively. Aligned muCP was used to direct the growth of RGC neurites from pure cultures (n=105) and retinal explants (n=64) along laminin patterns and to individual microelectrodes. Immunofluorescence stains (n=39) were used to determine the cell types that grew out from the retinal explants.


RESULTS
RGC neurite growth was directed reproducibly along aligned laminin micropatterns to individual microelectrodes in pure RGC cultures and from full-thickness explanted rat retinas in 92% of experiments, neurites from pure RGC cultures extended along the laminin lines with an average length of 263 +/- 118 microm (SD; n=27) after 24 hours. Neurites from retinal explants extended in more than 80% of experiments and were observed to grow to an average length of 279 +/- 78 microm (n=64) after 2 days in culture. These neurites grew up to 3 mm after 1 month of culture on the laminin micropatterns. Immunohistochemical stains demonstrated that extended processes from both RGCs and glial cells grew out of retinal explants onto stamped laminin lines.


CONCLUSIONS
Using muCP to pattern surfaces with growth factors, individual neuronal processes from pure RGC culture and whole retinal explants can be directed to discrete sites on a microelectronic chip surface. By directing RGC neurite processes to specific sites, single cell stimulation becomes possible. This may allow discrete populations of retinal neurons to be addressed so that physiologic retinal processing of visual information can be achieved.
Leng
Theodore
T
Ophthalmic Tissue Engineering Laboratory, Department of Ophthalmology, Stanford University School of Medicine, Stanford, California 94305, USA.
Wu
Peggy
P
Mehenti
Neville Z
NZ
Bent
Stacey F
SF
Marmor
Michael F
MF
Blumenkranz
Mark S
MS
Fishman
Harvey A
HA
D000818:Animals; D000831:Animals, Newborn; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004567:Electrodes, Implanted; D019084:Fluorescent Antibody Technique, Indirect; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D008839:Microelectrodes; D016501:Neurites / Q000502:physiology; D016900:Neurofilament Proteins / Q000378:metabolism; D019736:Prostheses and Implants; D051381:Rats; D017207:Rats, Sprague-Dawley; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000502:physiology; D018800:Thy-1 Antigens / Q000378:metabolism
D005904:Glial Fibrillary Acidic Protein; D016900:Neurofilament Proteins; D018800:Thy-1 Antigens; C051774:neurofilament protein H