Gaudin C, Forster V, Sahel J, Dreyfus H, Hicks D
Physiopathologie Rétinienne INSERM CJF-9202, Strasbourg, France.
Invest Ophthalmol Vis Sci. 1996 Oct;37(11):2258-68.
Fully mature neurons of central nervous system origin generally are considered unable to survive for extended periods of time in simple culture conditions. The authors report that adult and aged human, porcine, and rodent retinal neurons, including rod and cone photoreceptors, constitute an exception to this idea.
Cells were dissociated from human postmortem retinas, adult mammalian retinas, and selected brain regions and were seeded into tissue culture plates and left to develop as monolayer cultures for up to 2 months. A battery of antibody markers was used to identify the nature and morphology of the cells in vitro.
Photoreceptor cell survival of rods and cones was observed routinely when the delay between the time of death until culture preparation was 50 hours or less, compatible with current eye bank practice. Two-week-old cultures were formed of rod photoreceptors, representing approximately 50% of neuronal cell types; cone photoreceptors, representing 5% to 30% of neuronal cell types; other retinal neurons (especially amacrine cells approximately 20%); and retinal glial cells, present in variable numbers. Glial cells were essential for long-term photoreceptor survival and neurite outgrowth. Adult mammalian brain neurons isolated under the same conditions did not survive.
Fully adult human and other mammalian retinal neurons, including photoreceptors, exhibit remarkable plasticity in vitro, and such monolayer models may have applications in physiological, pharmacologic, and toxicologic studies of human and other mammalian retina.
中枢神经系统来源的完全成熟神经元通常被认为在简单培养条件下无法长时间存活。作者报告称,成人和老年人类、猪及啮齿动物的视网膜神经元,包括视杆和视锥光感受器,是这一观点的例外。
从人类死后视网膜、成年哺乳动物视网膜及选定的脑区分离细胞,接种到组织培养板中,作为单层培养物培养长达2个月。使用一系列抗体标记物来鉴定体外培养细胞的性质和形态。
当从死亡到培养准备的延迟时间为50小时或更短(符合当前眼库操作)时,常规观察到视杆和视锥光感受器细胞的存活。两周大的培养物由视杆光感受器组成,约占神经元细胞类型的50%;视锥光感受器,占神经元细胞类型的5%至30%;其他视网膜神经元(尤其是约20%的无长突细胞);以及数量不等的视网膜神经胶质细胞。神经胶质细胞对光感受器的长期存活和神经突生长至关重要。在相同条件下分离的成年哺乳动物脑神经元无法存活。
完全成熟的人类和其他哺乳动物视网膜神经元,包括光感受器,在体外表现出显著的可塑性,这种单层模型可能在人类和其他哺乳动物视网膜的生理、药理和毒理学研究中具有应用价值。