April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Time Course Of Organotypic Culture Degeneration Of Human Retina
Author Affiliations & Notes
  • Jose-Carlos Pastor
    IOBA-Campus Miguel Delibes, Fisiologia, Genetica y Microbiologia,
    University of Valladolid, Valladolid, Spain
  • Ivan Fernandez-Bueno
    IOBA-Campus Miguel Delibes, Fisiologia, Genetica y Microbiologia,
    University of Valladolid, Valladolid, Spain
  • Laura Fernandez-Sanchez
    IOBA-Campus Miguel Delibes, Fisiologia, Genetica y Microbiologia,
    Universidad de Alicante, Alicante, Spain
  • Maria Teresa Garcia-Gutierrez
    IOBA-Campus Miguel Delibes, Fisiologia, Genetica y Microbiologia,
    University of Valladolid, Valladolid, Spain
  • Manuel Gayoso
    Biologia Celular, Histologia y Farmacologia, Fisiologia Genetica y Microbiologia,
    University of Valladolid, Valladolid, Spain
  • Nicolas Cuenca
    Biologia Celular, Histologia y Farmacologia, Fisiologia Genetica y Microbiologia,
    Universidad de Alicante, Alicante, Spain
  • Footnotes
    Commercial Relationships  Jose-Carlos Pastor, None; Ivan Fernandez-Bueno, None; Laura Fernandez-Sanchez, None; Maria Teresa Garcia-Gutierrez, None; Manuel Gayoso, None; Nicolas Cuenca, None
  • Footnotes
    Support  MICINN (BFU2009-07793/BFI), (RETICS RD07/0062/0012) to NC. IF was supported by Junta de Castilla y León, Spain
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5366. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Jose-Carlos Pastor, Ivan Fernandez-Bueno, Laura Fernandez-Sanchez, Maria Teresa Garcia-Gutierrez, Manuel Gayoso, Nicolas Cuenca; Time Course Of Organotypic Culture Degeneration Of Human Retina. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5366.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : To characterise an ex-vivo adult human retina organ culture, to establish the survival pattern and early degeneration of neural and glial cells

Methods: : Two postmortem human globes from donors aged 57 and 59 years old were dissected and 16 neuroretina explants (5x5 mm) were obtained. Four were used as freshly retina controls and 12 were explanted on Transwell® culture dishes. Culture medium composed of Neurobasal A supplemented with B-27, FBS and L-glutamine was used. Cultured explants were kept for 3 (n=4), 6 (n=4) and 9 (n=4) days. Vertical epoxi resin sections, stained with toluidine blue, and cryostat sections, singly or doubly immunostained for specific markers of neuronal and glial cells, were used for evaluation.

Results: : Retinal morphology was properly preserved at culture start-point. Progressive retinal degeneration was observed, including early pyknosis of photoreceptor nuclei; cellular vacuolization in the ganglion cell layer; diminution of the plexiform layers thicknes; disruption and truncation of photoreceptor outer segments (OS); and marked reduction in the number of nuclei at both nuclear layers, where cells appeared less densely packed. Remarkable changes at 3 days included swelling of cone OS with impaired of their pedicles; loss of axon and dendrites of horizontal and rod bipolar cells stained with antibodies against calbindin (CB) and protein kinase C (PKC), respectively. At end-point (9 days), horizontal cells presented pyknotic nuclei and their terminal tips were gone. Similar degenerative processes were found in the outer plexiform layer for rod bipolar cells and a loss of axon terminal lateral varicosities in the inner plexiform layer. Glial fibrillary acidic protein (GFAP) staining did not reveal an increase of gliosis by Müller cells. However, some Müller cells were CB inmunoreactive since 6 days of culture

Conclusions: : These morphological changes, early observed at culture in cones, particularly in their OS and pedicles, and in their postsynaptic cells (horizontal and bipolar cells), no previously describe in human retina samples, indicate the important relationship of the retina with the pigment epithelium. These results may suggest the importance of time in retinal detachment treatment

Keywords: retinal culture • retinal detachment • immunohistochemistry 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×