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Immunocytochemical Profiling of Cultured Mouse Primary Retinal Cells

Zalis, Marina C. LU ; Johansson, Sebastian and Englund-Johansson, Ulrica LU (2017) In Journal of Histochemistry and Cytochemistry 65(4). p.223-239
Abstract

Primary retinal cell cultures and immunocytochemistry are important experimental platforms in ophthalmic research. Translation of retinal cells from their native environment to the in vitro milieu leads to cellular stress, jeopardizing their in vivo phenotype features. Moreover, the specificity and stability of many retinal immunochemical markers are poorly evaluated in retinal cell cultures. Hence, we here evaluated the expression profile of 17 retinal markers, that is, recoverin, rhodopsin, arrestin, Chx10, PKC, DCX, CRALBP, GS, vimentin, TPRV4, RBPMS, Brn3a, β-tubulin III, NeuN, MAP2, GFAP, and synaptophysin. At 7 and 18 days of culture, the marker expression profiles of mouse postnatal retinal cells were compared with their... (More)

Primary retinal cell cultures and immunocytochemistry are important experimental platforms in ophthalmic research. Translation of retinal cells from their native environment to the in vitro milieu leads to cellular stress, jeopardizing their in vivo phenotype features. Moreover, the specificity and stability of many retinal immunochemical markers are poorly evaluated in retinal cell cultures. Hence, we here evaluated the expression profile of 17 retinal markers, that is, recoverin, rhodopsin, arrestin, Chx10, PKC, DCX, CRALBP, GS, vimentin, TPRV4, RBPMS, Brn3a, β-tubulin III, NeuN, MAP2, GFAP, and synaptophysin. At 7 and 18 days of culture, the marker expression profiles of mouse postnatal retinal cells were compared with their age-matched in vivo retinas. We demonstrate stable in vitro expression of all markers, except for arrestin and CRALBP. Differences in cellular expression and location of some markers were observed, both over time in culture and compared with the age-matched retina. We hypothesize that these differences are likely culture condition dependent. Taken together, we suggest a thorough evaluation of the antibodies in specific culture settings, before extrapolating the in vitro results to an in vivo setting. Moreover, the identification of specific cell types may require a combination of different genes expressed or markers with structural information.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
glia, immunocytochemistry, immunohistochemistry, photoreceptors, primary retinal cell cultures, retinal markers, RGCs
in
Journal of Histochemistry and Cytochemistry
volume
65
issue
4
pages
17 pages
publisher
Histochemical Society
external identifiers
  • scopus:85018769665
  • wos:000400160100003
ISSN
0022-1554
DOI
10.1369/0022155416689675
language
English
LU publication?
yes
id
0ca58553-47b5-4d68-8277-8d6c9332c0ac
date added to LUP
2017-06-14 09:10:39
date last changed
2018-01-07 12:07:39
@article{0ca58553-47b5-4d68-8277-8d6c9332c0ac,
  abstract     = {<p>Primary retinal cell cultures and immunocytochemistry are important experimental platforms in ophthalmic research. Translation of retinal cells from their native environment to the in vitro milieu leads to cellular stress, jeopardizing their in vivo phenotype features. Moreover, the specificity and stability of many retinal immunochemical markers are poorly evaluated in retinal cell cultures. Hence, we here evaluated the expression profile of 17 retinal markers, that is, recoverin, rhodopsin, arrestin, Chx10, PKC, DCX, CRALBP, GS, vimentin, TPRV4, RBPMS, Brn3a, β-tubulin III, NeuN, MAP2, GFAP, and synaptophysin. At 7 and 18 days of culture, the marker expression profiles of mouse postnatal retinal cells were compared with their age-matched in vivo retinas. We demonstrate stable in vitro expression of all markers, except for arrestin and CRALBP. Differences in cellular expression and location of some markers were observed, both over time in culture and compared with the age-matched retina. We hypothesize that these differences are likely culture condition dependent. Taken together, we suggest a thorough evaluation of the antibodies in specific culture settings, before extrapolating the in vitro results to an in vivo setting. Moreover, the identification of specific cell types may require a combination of different genes expressed or markers with structural information.</p>},
  author       = {Zalis, Marina C. and Johansson, Sebastian and Englund-Johansson, Ulrica},
  issn         = {0022-1554},
  keyword      = {glia,immunocytochemistry,immunohistochemistry,photoreceptors,primary retinal cell cultures,retinal markers,RGCs},
  language     = {eng},
  month        = {04},
  number       = {4},
  pages        = {223--239},
  publisher    = {Histochemical Society},
  series       = {Journal of Histochemistry and Cytochemistry},
  title        = {Immunocytochemical Profiling of Cultured Mouse Primary Retinal Cells},
  url          = {http://dx.doi.org/10.1369/0022155416689675},
  volume       = {65},
  year         = {2017},
}