Experimental Eye Research Institute, University Eye Hospital, 9142Ruhr-University Bochum, Germany.
Centre for Ophthalmology Tübingen, 27203University Eye Hospital Tübingen, Germany.
Altern Lab Anim. 2022 Jan;50(1):27-44. doi: 10.1177/02611929221082662. Epub 2022 Mar 18.
Neural retinal organ cultures are used to investigate ocular pathomechanisms. However, these cultures lack the essential retinal pigment epithelium (RPE) cells, which are part of the actual retina. To simulate a more realistic model, porcine neural retina explants were cocultured with ARPE-19 cells (ARPE-19 group), which are derived from human RPE. To identify whether the entire cells or just the cell factors are necessary, in a second experimental group, porcine neural retina explants were cultured with medium derived from ARPE-19 cells (medium group). Individually cultured neural retina explants served as controls (control group). After 8 days, all neural retinas were analysed to evaluate retinal thickness, photoreceptors, microglia, complement factors and synapses ( = 6-8 per group). The neural retina thickness in the ARPE-19 group was significantly better preserved than in the control group ( = 0.031). Also, the number of L-cones was higher in the ARPE-19 group, as compared to the control group ( < 0.001). Furthermore, the ARPE-19 group displayed an increased presynaptic glutamate uptake (determined via vGluT1 labelling) and enhanced post-synaptic density (determined via PSD-95 labelling). Combined Iba1 and iNOS detection revealed only minor effects of ARPE-19 cells on microglial activity, with a slight downregulation of total microglia activity apparent in the medium group. Likewise, only minor beneficial effects on photoreceptors and synaptic structure were found in the medium group. This novel system offers the opportunity to investigate interactions between the neural retina and RPE cells, and suggests that the inclusion of a RPE feeder layer has beneficial effects on the maintenance of neural retina. By modifying the culture conditions, this coculture model allows a better understanding of photoreceptor death and photoreceptor-RPE cell interactions in retinal diseases.
神经视网膜器官培养用于研究眼部发病机制。然而，这些培养物缺乏视网膜色素上皮 (RPE) 细胞，而后者是实际视网膜的一部分。为了模拟更逼真的模型，将猪的神经视网膜外植体与 ARPE-19 细胞（ARPE-19 组）共培养，ARPE-19 细胞源自人 RPE。为了确定是整个细胞还是仅仅是细胞因子是必需的，在第二个实验组中，将猪的神经视网膜外植体用源自 ARPE-19 细胞的培养基（培养基组）进行培养。单独培养的神经视网膜外植体作为对照（对照组）。培养 8 天后，所有神经视网膜均进行分析以评估视网膜厚度、光感受器、小胶质细胞、补体因子和突触（每组 6-8 个）。ARPE-19 组的神经视网膜厚度明显优于对照组（ = 0.031）。此外，与对照组相比，ARPE-19 组的 L-锥体数量更多（ < 0.001）。此外，ARPE-19 组显示出增加的突触前谷氨酸摄取（通过 vGluT1 标记测定）和增强的突触后密度（通过 PSD-95 标记测定）。Iba1 和 iNOS 的联合检测仅显示 ARPE-19 细胞对小胶质细胞活性的微小影响，培养基组中总小胶质细胞活性明显下调。同样，在培养基组中仅发现对光感受器和突触结构的微小有益影响。这种新系统提供了研究神经视网膜和 RPE 细胞之间相互作用的机会，并表明包含 RPE 饲养层对维持神经视网膜具有有益作用。通过改变培养条件，这种共培养模型可以更好地理解视网膜疾病中光感受器死亡和光感受器-RPE 细胞相互作用。
