Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension

Peter D. Westenskow, Toshihide Kurihara, Stephen Bravo, Daniel Feitelberg, Zack A. Sedillo, Edith Aguilar, Martin Friedlander

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

Original languageEnglish
Article numbere52247
JournalJournal of Visualized Experiments
Issue number95
DOIs
Publication statusPublished - 2015 Jan 23

Fingerprint

Retinal Pigments
Retinal Pigment Epithelium
Pigments
Rodentia
Suspensions
Injections
Macular Degeneration
Stem cells
Needles
Animals
Animal Models
Retinal Cone Photoreceptor Cells
Retinal Rod Photoreceptor Cells
Syringes
Pluripotent Stem Cells
Vertebrate Photoreceptor Cells
Eye Diseases
Touch
Retina
Cones

Keywords

  • Age-related macular degeneration
  • Cell-based delivery
  • Issue 95
  • Medicine
  • Retinal pigment epithelium
  • Subretinal injections
  • Translational medicine

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Westenskow, P. D., Kurihara, T., Bravo, S., Feitelberg, D., Sedillo, Z. A., Aguilar, E., & Friedlander, M. (2015). Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension. Journal of Visualized Experiments, (95), [e52247]. https://doi.org/10.3791/52247

Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension. / Westenskow, Peter D.; Kurihara, Toshihide; Bravo, Stephen; Feitelberg, Daniel; Sedillo, Zack A.; Aguilar, Edith; Friedlander, Martin.

In: Journal of Visualized Experiments, No. 95, e52247, 23.01.2015.

Research output: Contribution to journalArticle

Westenskow, PD, Kurihara, T, Bravo, S, Feitelberg, D, Sedillo, ZA, Aguilar, E & Friedlander, M 2015, 'Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension', Journal of Visualized Experiments, no. 95, e52247. https://doi.org/10.3791/52247
Westenskow PD, Kurihara T, Bravo S, Feitelberg D, Sedillo ZA, Aguilar E et al. Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension. Journal of Visualized Experiments. 2015 Jan 23;(95). e52247. https://doi.org/10.3791/52247
Westenskow, Peter D. ; Kurihara, Toshihide ; Bravo, Stephen ; Feitelberg, Daniel ; Sedillo, Zack A. ; Aguilar, Edith ; Friedlander, Martin. / Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension. In: Journal of Visualized Experiments. 2015 ; No. 95.
@article{36444615e02d479599a99c6331d4d27b,
title = "Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension",
abstract = "The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.",
keywords = "Age-related macular degeneration, Cell-based delivery, Issue 95, Medicine, Retinal pigment epithelium, Subretinal injections, Translational medicine",
author = "Westenskow, {Peter D.} and Toshihide Kurihara and Stephen Bravo and Daniel Feitelberg and Sedillo, {Zack A.} and Edith Aguilar and Martin Friedlander",
year = "2015",
month = "1",
day = "23",
doi = "10.3791/52247",
language = "English",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "95",

}

TY - JOUR

T1 - Performing subretinal injections in rodents to deliver retinal pigment epithelium cells in suspension

AU - Westenskow, Peter D.

AU - Kurihara, Toshihide

AU - Bravo, Stephen

AU - Feitelberg, Daniel

AU - Sedillo, Zack A.

AU - Aguilar, Edith

AU - Friedlander, Martin

PY - 2015/1/23

Y1 - 2015/1/23

N2 - The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

AB - The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

KW - Age-related macular degeneration

KW - Cell-based delivery

KW - Issue 95

KW - Medicine

KW - Retinal pigment epithelium

KW - Subretinal injections

KW - Translational medicine

UR - http://www.scopus.com/inward/record.url?scp=84923585981&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84923585981&partnerID=8YFLogxK

U2 - 10.3791/52247

DO - 10.3791/52247

M3 - Article

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 95

M1 - e52247

ER -