Transplantation of cultured adult porcine full-thickness retina
Transplantation of cultured adult porcine full-thickness retina
复制标题
DOI：
10.3727/000000007783464506
复制
发表时间：
2007-01-01
影响因子：
3.3
通讯作者：
Ghosh, Fredrik
中科院分区：
文献类型：
--
作者：
Engelsberg, Karl;Ghosh, Fredrik
在这项研究中，我们想研究一只眼睛与人类相似的动物的成年神经视网膜如何在体外存活。我们还想研究当用于移植范例时，培养过程如何影响成人视网膜。将来自成年猪眼的全层神经视网膜片解剖成直径为3 mm的片。将其保持在培养物中1-3天。此后，将外植体固定或视网膜下移植至成年猪，72-74天后处死。对移植的眼睛以及仅保持在培养物中的组织进行苏木精和伊红染色和免疫组织化学处理。外植体在离体培养（DIV）中保持1天显示出正常的形态。在这些标本中，单个固缩细胞在外核层（ONL）和神经节细胞层中明显，但更常见于内核层（INL）。在体外培养较长时间后，出现严重的退行性变化。移植前保持IDIV的移植外植体在四个样本中的两个中表现出正常的视网膜分层。Transducin和recoverin标记显示，在这些移植物的光感受器的内部段。视杆双极细胞形态正常。垂直排列的Muller细胞也见于层压移植物中。三个移植物中的两个保持2 DIV显示最小分层。移植眼在移植前保持3DIV显示所有眼的移植物变性。这项研究表明，成年猪神经视网膜外植体保持在培养1天显示正常的形态在其主要部分。此外，即使在几个月后，1天外植体也可以在保留形态的情况下存活移植。这表明在收获和移植之间储存成人供体组织的可能性。该培养系统在将来也可以用作在移植之前操纵视网膜供体组织的工具。
In this study we wanted to examine how an adult neuroretina from an animal with an eye similar to the human one survives in vitro. We also wanted to investigate how the culture process affects the adult retina when used in a transplantation paradigm. Full-thickness neuroretinal sheets from adult porcine eyes were dissected into pieces measuring 3 mm in diameter. These were kept in culture for 1-3 days. After this time, the explants were fixed or transplanted subretinally to adult pigs, which were killed after 72-74 days. Transplanted eyes, as well as tissue kept in culture only, were processed for hematoxylin and eosin staining and immunohistochemistry. Explants kept I day in vitro (DIV) displayed the normal morphology. In these specimens, single pyknotic cells were evident in the outer nuclear layer (ONL) and ganglion cell layer, but were more frequent in the inner nuclear layer (INL). After longer times in vitro, severe degenerative changes appeared. Transplanted explants kept I DIV prior to transplantation exhibited normal retina] lamination in two out of four specimens. Transducin and recoverin labeling revealed photoreceptors with inner segments in these grafts. Rod bipolar cells displayed a normal morphology. Vertically arranged Muller cells were also seen in the laminated grafts. Two of the three transplants kept 2 DIV displayed minimal lamination. Eyes with transplants kept 3 DIV prior to transplantation displayed degenerated grafts in all eyes. This study shows that adult porcine neuroretinal explants kept in culture for I day display a normal morphology in their major part. Additionally, 1-day explants can survive transplantation with retained morphology even after several months. This indicates the possibility of storing adult donor tissue between harvest and transplantation. The culture system may also be used in the future as a tool for manipulating retina] donor tissue prior to transplantation.