gms | German Medical Science

25th Annual Meeting of the German Retina Society

German Retina Society

01.06. - 02.06.2012, Münster

Establishment of a retinal ischemia organ culture model

Meeting Abstract

  • presenting/speaker Sven Schnichels - Universitäts-Augenklinik Tübingen
  • M. Blak - Universitäts-Augenklinik Tübingen
  • J. Hofmann - Universitäts-Augenklinik Tübingen
  • K.U. Bartz-Schmidt - Universitäts-Augenklinik Tübingen
  • M.S. Spitzer - Universitäts-Augenklinik Tübingen
  • M. Schultheiss - Universitäts-Augenklinik Tübingen

German Retina Society. 25th Annual Conference of the German Retina Society. Münster, 01.-02.06.2012. Düsseldorf: German Medical Science GMS Publishing House; 2012. Doc12rg09

DOI: 10.3205/12rg09, URN: urn:nbn:de:0183-12rg097

This is the translated version of the article.
The original version can be found at: http://www.egms.de/de/meetings/rg2012/12rg09.shtml

Published: May 30, 2012

© 2012 Schnichels et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.


Outline

Text

Background: Ischemia plays an important role in several ophthalmological diseases. Retinal ganglion cells (RGCs) are the most sensitive cells to ischemia. To investigate neuroprotective agents and therapies against these ophthalmological diseases we developed an easy-to-use chamber for organotypic cultures. We decided to use organotypic cultures, because in-vivo models or primary cultures are very time-consuming, expensive and several therapies or agents can not be tested in these models.

Methods: For this pilot-study, we incubated retinas at 37°C for five different time periods (45–120 minutes) under ischemic conditions. The chamber was streamed with N2 for 5 minutes, then the chamber was immediately sealed and the retinas were incubated for the rest of the designated time. Thereafter the organotypic cultures were incubated for 24, 48 or 72 h in an incubator under standard conditions. To analyze the amount of RGCs and apoptotic RGCs, the retinas were frozen and processed for cutting. RGCs immunohistology was performed with a Brna3a-antibody. Apoptotic cells were visualized via TUNEL-staining and overall cell amount via DAPI-staining.

Results: A time-dependant decrease in the amount of RGCs after 24, 48 or 72 h was observable. Moreover, also a decrease in the amount of RGCs that was dependant on ischemia duration was observed. Furthermore the amount of TUNEL-positive cells was ischemia dependant.

Conclusion: We successfully established an organotypic culture model for retinal ischemia. Any therapy can now be tested under ischemic organotypic conditions. Furthermore, we improved our chamber to allow manipulations (injection of drugs, exchange of the media or temperature adjustment) during ischemia.