Development of the embryonic porcine neuroretina in vitro.
OBJECTIVE
The objective of this study was to investigate the survival and morphology of embryonic porcine full-thickness neuroretina in culture.


METHODS
Porcine fetuses were taken out by cesarian section, and the eyes were enucleated. Neuroretinas were explanted on culture plate inserts and were kept for 0-42 days in vitro under standard culture conditions. Green nucleic acid (Sytox) was used for measuring the extent of cell death, and 4,6-diaminidine-2-phenylindoldihydrochloride was used as a marker for the cellular layers. The explants were examined as whole-mount preparations and vertical sections. Sectioned tissue was stained with hematoxylin-eosin and labeled for immunohistochemistry with photoreceptor-specific antibodies raised against transducin and recoverin.


RESULTS
In explants kept for 0-5 days in vitro, the developing retina consisted of multiple rows of neuroblastic cells and a more defined, but multilayered ganglion cell layer (GCL). Older explants revealed a more differentiated appearance with ultimately all normal retinal layers present, even after 42 days in vitro. Transducin- and recoverin-labeled photoreceptors were seen in these specimens, but no outer segments were found. The whole-mount preparation revealed extensively Sytox-labeled cells in the GCL at 2 days in vitro, but very few cells were labeled in older explants.


CONCLUSIONS
This study shows that cultured fetal porcine full-thickness neuroretina can survive and develop according to its intrinsic timetable for at least 6 weeks in vitro. The in vitro system for culturing of the full-thickness retina may be useful in experiments involving retinal transplantation.
Engelsberg
Karl
K
Department of Ophthalmology, University Hospital, SE-22185 Lund, Sweden. karl.engelsberg@oft.lu.se
Johansson
Kjell
K
Ghosh
Fredrik
F
D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002470:Cell Survival; D005136:Eye Proteins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D007150:Immunohistochemistry; D007211:Indoles / Q000378:metabolism; D008074:Lipoproteins / Q000378:metabolism; D009924:Organ Culture Techniques; D009930:Organic Chemicals; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D051603:Recoverin; D012160:Retina / Q000196:embryology* / Q000378:metabolism; D013552:Swine / Q000033:anatomy & histology*; D015293:Transducin / Q000378:metabolism
D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D005456:Fluorescent Dyes; D007211:Indoles; D008074:Lipoproteins; D009930:Organic Chemicals; C402795:SYTOX Green; D051603:Recoverin; C007293:DAPI; D015293:Transducin