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Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology

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Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology. / Alarautalahti, Virpi; Ragauskas, Symantas; Hakkarainen, Jenni J.; Uusitalo-Järvinen, Hannele; Uusitalo, Hannu; Hyttinen, Jari; Kalesnykas, Giedrius; Nymark, Soile.

In: Investigative ophthalmology & visual science, Vol. 60, No. 6, 01.05.2019, p. 1914-1927.

Research output: Contribution to journalArticleScientificpeer-review

Harvard

Alarautalahti, V, Ragauskas, S, Hakkarainen, JJ, Uusitalo-Järvinen, H, Uusitalo, H, Hyttinen, J, Kalesnykas, G & Nymark, S 2019, 'Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology', Investigative ophthalmology & visual science, vol. 60, no. 6, pp. 1914-1927. https://doi.org/10.1167/iovs.18-25156

APA

Alarautalahti, V., Ragauskas, S., Hakkarainen, J. J., Uusitalo-Järvinen, H., Uusitalo, H., Hyttinen, J., ... Nymark, S. (2019). Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology. Investigative ophthalmology & visual science, 60(6), 1914-1927. https://doi.org/10.1167/iovs.18-25156

Vancouver

Alarautalahti V, Ragauskas S, Hakkarainen JJ, Uusitalo-Järvinen H, Uusitalo H, Hyttinen J et al. Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology. Investigative ophthalmology & visual science. 2019 May 1;60(6):1914-1927. https://doi.org/10.1167/iovs.18-25156

Author

Alarautalahti, Virpi ; Ragauskas, Symantas ; Hakkarainen, Jenni J. ; Uusitalo-Järvinen, Hannele ; Uusitalo, Hannu ; Hyttinen, Jari ; Kalesnykas, Giedrius ; Nymark, Soile. / Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology. In: Investigative ophthalmology & visual science. 2019 ; Vol. 60, No. 6. pp. 1914-1927.

Bibtex - Download

@article{8593ee33ae1e4bf596ea76bd7c5676d8,
title = "Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology",
abstract = "Purpose: Retinal explant cultures provide simplified systems where the functions of the retina and the effects of ocular therapies can be studied in an isolated environment. The purpose of this study was to provide insight into long-term preservation of retinal tissue in culture conditions, enable a deeper understanding of the interdependence of retinal morphology and function, and ensure the reliability of the explant technique for prolonged experiments. Methods: Retinal explants from adult mice were cultured as organotypic culture at the air-medium interface for 14 days in vitro (DIV). Retinal functionality was assessed by multielectrode array technique and morphology by immunohistochemical methods at several time points during culture. Results: Retinal explants retained viability for 14 DIV, although with diminishing neuronal activity, progressing neuronal loss, and increasing reactive gliosis. We recorded spontaneous retinal ganglion cell (RGC) activity up to 14 DIV with temporally changing distribution of RGC firing rates. Light responsiveness was measurable from RGCs for 7 DIV and from photoreceptors for 2 DIV. Apoptotic cells were detected beginning at 3 DIV with their density peaking at 7 DIV. The number of RGCs gradually decreased by 70{\%} during 14 DIV. The change was accompanied by the loss of RGC functionality, resulting in 84{\%} loss of electrically active RGCs. Conclusions: Retinal explants provide a valuable tool for studies of retinal functions and development of ocular therapies. However, critical for long-term use, retinal functionality was lost before structural loss, emphasizing a need for both functional and morphologic readouts to determine the overall state of the cultured retina.",
author = "Virpi Alarautalahti and Symantas Ragauskas and Hakkarainen, {Jenni J.} and Hannele Uusitalo-J{\"a}rvinen and Hannu Uusitalo and Jari Hyttinen and Giedrius Kalesnykas and Soile Nymark",
note = "DUPL=47898722",
year = "2019",
month = "5",
day = "1",
doi = "10.1167/iovs.18-25156",
language = "English",
volume = "60",
pages = "1914--1927",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology",
number = "6",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Viability of Mouse Retinal Explant Cultures Assessed by Preservation of Functionality and Morphology

AU - Alarautalahti, Virpi

AU - Ragauskas, Symantas

AU - Hakkarainen, Jenni J.

AU - Uusitalo-Järvinen, Hannele

AU - Uusitalo, Hannu

AU - Hyttinen, Jari

AU - Kalesnykas, Giedrius

AU - Nymark, Soile

N1 - DUPL=47898722

PY - 2019/5/1

Y1 - 2019/5/1

N2 - Purpose: Retinal explant cultures provide simplified systems where the functions of the retina and the effects of ocular therapies can be studied in an isolated environment. The purpose of this study was to provide insight into long-term preservation of retinal tissue in culture conditions, enable a deeper understanding of the interdependence of retinal morphology and function, and ensure the reliability of the explant technique for prolonged experiments. Methods: Retinal explants from adult mice were cultured as organotypic culture at the air-medium interface for 14 days in vitro (DIV). Retinal functionality was assessed by multielectrode array technique and morphology by immunohistochemical methods at several time points during culture. Results: Retinal explants retained viability for 14 DIV, although with diminishing neuronal activity, progressing neuronal loss, and increasing reactive gliosis. We recorded spontaneous retinal ganglion cell (RGC) activity up to 14 DIV with temporally changing distribution of RGC firing rates. Light responsiveness was measurable from RGCs for 7 DIV and from photoreceptors for 2 DIV. Apoptotic cells were detected beginning at 3 DIV with their density peaking at 7 DIV. The number of RGCs gradually decreased by 70% during 14 DIV. The change was accompanied by the loss of RGC functionality, resulting in 84% loss of electrically active RGCs. Conclusions: Retinal explants provide a valuable tool for studies of retinal functions and development of ocular therapies. However, critical for long-term use, retinal functionality was lost before structural loss, emphasizing a need for both functional and morphologic readouts to determine the overall state of the cultured retina.

AB - Purpose: Retinal explant cultures provide simplified systems where the functions of the retina and the effects of ocular therapies can be studied in an isolated environment. The purpose of this study was to provide insight into long-term preservation of retinal tissue in culture conditions, enable a deeper understanding of the interdependence of retinal morphology and function, and ensure the reliability of the explant technique for prolonged experiments. Methods: Retinal explants from adult mice were cultured as organotypic culture at the air-medium interface for 14 days in vitro (DIV). Retinal functionality was assessed by multielectrode array technique and morphology by immunohistochemical methods at several time points during culture. Results: Retinal explants retained viability for 14 DIV, although with diminishing neuronal activity, progressing neuronal loss, and increasing reactive gliosis. We recorded spontaneous retinal ganglion cell (RGC) activity up to 14 DIV with temporally changing distribution of RGC firing rates. Light responsiveness was measurable from RGCs for 7 DIV and from photoreceptors for 2 DIV. Apoptotic cells were detected beginning at 3 DIV with their density peaking at 7 DIV. The number of RGCs gradually decreased by 70% during 14 DIV. The change was accompanied by the loss of RGC functionality, resulting in 84% loss of electrically active RGCs. Conclusions: Retinal explants provide a valuable tool for studies of retinal functions and development of ocular therapies. However, critical for long-term use, retinal functionality was lost before structural loss, emphasizing a need for both functional and morphologic readouts to determine the overall state of the cultured retina.

U2 - 10.1167/iovs.18-25156

DO - 10.1167/iovs.18-25156

M3 - Article

VL - 60

SP - 1914

EP - 1927

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 6

ER -