Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry

Sumana R. Chintalapudi, Need N. Patel, Zachary K. Goldsmith, Levon Djenderedjian, Xiang D. Di Wang, Tony Marion, Monica Jablonski, Vanessa Morales-Tirado

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.

Original languageEnglish (US)
Article numbere55785
JournalJournal of Visualized Experiments
Volume2017
Issue number125
DOIs
StatePublished - Jul 5 2017

Fingerprint

Neurodegenerative diseases
Flow cytometry
Retinal Ganglion Cells
Flow Cytometry
Aging of materials
Neurodegenerative Diseases
Retina
Diabetic Retinopathy
Glaucoma
Patient Selection
Visual Acuity

All Science Journal Classification (ASJC) codes

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

Cite this

Chintalapudi, S. R., Patel, N. N., Goldsmith, Z. K., Djenderedjian, L., Di Wang, X. D., Marion, T., ... Morales-Tirado, V. (2017). Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry. Journal of Visualized Experiments, 2017(125), [e55785]. https://doi.org/10.3791/55785

Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry. / Chintalapudi, Sumana R.; Patel, Need N.; Goldsmith, Zachary K.; Djenderedjian, Levon; Di Wang, Xiang D.; Marion, Tony; Jablonski, Monica; Morales-Tirado, Vanessa.

In: Journal of Visualized Experiments, Vol. 2017, No. 125, e55785, 05.07.2017.

Research output: Contribution to journalArticle

Chintalapudi SR, Patel NN, Goldsmith ZK, Djenderedjian L, Di Wang XD, Marion T et al. Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry. Journal of Visualized Experiments. 2017 Jul 5;2017(125). e55785. https://doi.org/10.3791/55785
Chintalapudi, Sumana R. ; Patel, Need N. ; Goldsmith, Zachary K. ; Djenderedjian, Levon ; Di Wang, Xiang D. ; Marion, Tony ; Jablonski, Monica ; Morales-Tirado, Vanessa. / Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry. In: Journal of Visualized Experiments. 2017 ; Vol. 2017, No. 125.
@article{12b8725d5c184d8b8ecdd321c21fe97f,
title = "Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry",
abstract = "Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.",
author = "Chintalapudi, {Sumana R.} and Patel, {Need N.} and Goldsmith, {Zachary K.} and Levon Djenderedjian and {Di Wang}, {Xiang D.} and Tony Marion and Monica Jablonski and Vanessa Morales-Tirado",
year = "2017",
month = "7",
day = "5",
doi = "10.3791/55785",
language = "English (US)",
volume = "2017",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "125",

}

TY - JOUR

T1 - Isolation of primary murine retinal ganglion cells (RGCs) by flow cytometry

AU - Chintalapudi, Sumana R.

AU - Patel, Need N.

AU - Goldsmith, Zachary K.

AU - Djenderedjian, Levon

AU - Di Wang, Xiang D.

AU - Marion, Tony

AU - Jablonski, Monica

AU - Morales-Tirado, Vanessa

PY - 2017/7/5

Y1 - 2017/7/5

N2 - Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.

AB - Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.

UR - http://www.scopus.com/inward/record.url?scp=85024370774&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85024370774&partnerID=8YFLogxK

U2 - 10.3791/55785

DO - 10.3791/55785

M3 - Article

VL - 2017

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 125

M1 - e55785

ER -