Development of the embryonic porcine neuroretina in vitro

被引:20
作者
Engelsberg, K [1 ]
Johansson, K [1 ]
Ghosh, F [1 ]
机构
[1] Univ Lund Hosp, Dept Ophthalmol, SE-22185 Lund, Sweden
关键词
pig neuroretina; retinal development; in vitro; transducin; recoverin;
D O I
10.1159/000084252
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
Purpose: 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. Conclusion: 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. Copyright (C) 2005 S. Karger AG, Basel.
引用
收藏
页码:104 / 111
页数:8
相关论文
共 27 条
[1]   Membranes of retinal pigment epithelial cells in vitro are damaged in the phagocytotic process of the photoreceptor outer segment discs peroxidized by ferrous ions [J].
Akeo, K ;
Hiramitsu, T ;
Yorifuji, H ;
Okisaka, S .
PIGMENT CELL RESEARCH, 2002, 15 (05) :341-347
[2]  
ALTSHULER D, 1993, DEVELOPMENT, V119, P1317
[3]   TRANSPLANTATION OF EMBRYONIC RETINA TO THE SUBRETINAL SPACE IN RABBITS [J].
BERGSTROM, A ;
EHINGER, B ;
WILKE, K ;
ZUCKER, CL ;
ADOLPH, AR ;
ARAMANT, R ;
SEILER, M .
EXPERIMENTAL EYE RESEARCH, 1992, 55 (01) :29-37
[4]   HISTOTYPIC DIFFERENTIATION OF NEONATAL MOUSE RETINA IN ORGAN-CULTURE [J].
CAFFE, AR ;
VISSER, H ;
JANSEN, HG ;
SANYAL, S .
CURRENT EYE RESEARCH, 1989, 8 (10) :1083-1092
[5]   Cell death in the inner nuclear layer of the retina is modulated by BDNF [J].
Cusato, K ;
Bosco, A ;
Linden, R ;
Reese, BE .
DEVELOPMENTAL BRAIN RESEARCH, 2002, 139 (02) :325-330
[6]   Apoptotic cell death and microglial cell responses in cultured rat retina [J].
Engelsberg, K ;
Ehinger, B ;
Wassélius, J ;
Johansson, K .
GRAEFES ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY, 2004, 242 (03) :229-239
[7]  
García M, 2002, INVEST OPHTH VIS SCI, V43, P3735
[8]   APOPTOSIS IN ADULT RETINAL GANGLION-CELLS AFTER AXOTOMY [J].
GARCIAVALENZUELA, E ;
GORCZYCA, W ;
DARZYNKIEWICZ, Z ;
SHARMA, SC .
JOURNAL OF NEUROBIOLOGY, 1994, 25 (04) :431-438
[9]   Transplantation of full-thickness retina in the rhodopsin transgenic pig [J].
Ghosh, F ;
Wong, F ;
Johansson, K ;
Bruun, A ;
Peters, RM .
RETINA-THE JOURNAL OF RETINAL AND VITREOUS DISEASES, 2004, 24 (01) :98-109
[10]   Transplantation of full-thickness retina in the normal porcine eye -: Surgical and morphologic aspects [J].
Ghosh, F ;
Arnér, K .
RETINA-THE JOURNAL OF RETINAL AND VITREOUS DISEASES, 2002, 22 (04) :478-486