Functional and morphological analysis of the subretinal injection of retinal pigment epithelium cells.
Functional and morphological analysis of the subretinal injection of retinal pigment epithelium cells.
复制标题
DOI：
10.1017/s0952523812000041
复制
发表时间：
2012-03
影响因子：
1.9
通讯作者：
Williams, David S.
中科院分区：
文献类型：
--
作者：
Engelhardt, Maren;Tosha, Chinatsu;Lopes, Vanda S.;Chen, Bryan;Nguyen, Lisa;Nusinowitz, Steven;Williams, David S.
关键词：
视网膜色素上皮（RPE）细胞移植是一种潜在的治疗视网膜变性。在这里，我们使用侵入性和非侵入性方法相结合来表征视网膜下注射RPE细胞的结构和功能后果。将来自原代培养物的色素细胞注射到白化病小鼠中。通过眼底成像、光谱域光学相干断层扫描（sdOCT）、组织学和视网膜电图（ERG）在8周内监测恢复情况。sdOCT显示视网膜复位在1周时几乎完成。视网膜电图反应幅度减少后，注射，锥介导的功能，然后恢复优于杆功能。通过sdOCT和组织学观察，注射部位附近的感光细胞损失明显，可能是恢复不完全的主要原因。通过显微镜检查，通过顶端黑素体的存在来鉴定注射的细胞。它们要么与布鲁赫膜建立接触，从而成为RPE单层的一部分，要么位于宿主细胞的顶面，导致注射细胞的基底面与宿主细胞的顶面并置，并形成一系列桥粒连接。RPE细胞密度没有增加，表明注射细胞掺入RPE单层中伴随着宿主细胞的损失。移植的和剩余的宿主细胞含有摄入碎片的大空泡以及脂褐素样颗粒，表明它们已经清除了过量的注射细胞和宿主细胞，并受到高消化负荷的应激。因此，尽管观察到显著的功能和结构恢复，但这种消化应激的后果可能是长期健康的问题，特别是在RPE细胞移植用于治疗包括脂褐素积累作为其病理学一部分的疾病的情况下。
Replacement of retinal pigment epithelium (RPE) cells by transplantation is a potential treatment for some retinal degenerations. Here, we used a combination of invasive and noninvasive methods to characterize the structural and functional consequences of subretinal injection of RPE cells. Pigmented cells from primary cultures were injected into albino mice. Recovery was monitored over 8 weeks by fundus imaging, spectral domain optical coherence tomography (sdOCT), histology, and electroretinography (ERG). sdOCT showed that retinal reattachment was nearly complete by 1 week. ERG response amplitudes were reduced after injection, with cone-mediated function then recovering better than rod function. Photoreceptor cell loss was evident by sdOCT and histology, near the site of injection, and is likely to have been the main cause of incomplete recovery. With microscopy, injected cells were identified by the presence of apical melanosomes. They either established contact with Bruch’s membrane, and thus became part of the RPE monolayer, or were located on the apical surface of the host’s cells, resulting in apposition of the basal surface of the injected cell with the apical surface of the host cell and the formation of a series of desmosomal junctions. RPE cell density was not increased, indicating that the incorporation of an injected cell into the RPE monolayer was concomitant with the loss of a host cell. The transplanted and remaining host cells contained large vacuoles of ingested debris as well as lipofuscin-like granules, suggesting that they had scavenged the excess injected and host cells, and were stressed by the high digestive load. Therefore, although significant functional and structural recovery was observed, the consequences of this digestive stress may be a concern for longer-term health, especially where RPE cell transplantation is used to treat diseases that include lipofuscin accumulation as part of their pathology.