Adenovirally transduced bone marrow stromal cells differentiate into pigment epithelial cells and induce rescue effects in RCS rats
Adenovirally transduced bone marrow stromal cells differentiate into pigment epithelial cells and induce rescue effects in RCS rats
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DOI：
10.1167/iovs.04-1501
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发表时间：
2006-09-01
影响因子：
4.4
通讯作者：
Schraermeyer, Ulrich
中科院分区：
文献类型：
--
作者：
Arnhold, Stefan;Heiduschka, Peter;Schraermeyer, Ulrich
目的。目的：探讨腺病毒转染的骨髓基质细胞（BMSCs）向视网膜色素上皮样细胞分化的潜力，并评价BMSCs移植到英国皇家外科学院（RCS）大鼠视网膜后可能的挽救作用。通过表达绿色荧光蛋白（GFP）或色素上皮衍生因子（PEDF）的高容量腺病毒载体，在视网膜下移植前将大鼠间充质干细胞体外转导到Wistar大鼠或RCS大鼠体内。细胞注射两个月后，处死大鼠，去眼。然后对眼睛进行光学显微镜检查或电子显微镜检查。利用细胞角蛋白抗体和紧密连接蛋白ZO-1抗体对细胞分化和整合进行免疫细胞化学分析。注射细胞16天后进行视网膜电图检查是否能检测到功能性抢救。体外共培养人MSCs和人RPE细胞实验表明，MSCs具有RPE样特性。在移植实验中，部分大鼠间充质干细胞与Wistar和RCS大鼠宿主RPE细胞层结合，表现为六边形形态。视网膜下移植细胞表达上皮标志物细胞角蛋白，并与宿主RPE细胞建立紧密连接。此外，将载体转导的和非转导的间充质干细胞移植到营养不良视网膜的半薄切片后，可以证明有挽救作用。超微结构上，MSCs位于宿主RPE顶部，并与光感受器外段、吞噬杆外段紧密接触。综上所述，这些结果提高了MSCs具有替代病变RPE细胞并将治疗蛋白传递到视网膜下空间以保护光感受器细胞免受变性的可能性。
PURPOSE. To determine the potential of adenovirally transduced bone marrow stromal cells (BMSCs) to differentiate into retinal pigment epithelial-like cells and to evaluabe possible rescue effects after transplantation into the retinas of Royal College of Surgeons (RCS) rats.METHODS. Through a high-capacity adenoviral vector expressing either green fluorescent protein (GFP) or pigment epithelial-derived factor (PEDF), rat MSCs were transduced in vitro before subretinal transplantation into Wistar rats or, alternatively, RCS rats. Two months after cell injection, the rats were killed and the eyes enucleated. The eyes were then investigated light microscopically or processed for electron microscopic investigations. Cell differentiation and integration were analyzed immunocytochemically using antibodies against cytokeratin and the tight junction protein ZO-1. Electroretinography was performed 16 days after injection of cells, to check whether a functional rescue could be detected.RESULTS. In vitro experiments in cocultured human MSCs and human RPE cells showed that MSCs adopted RPE-like characteristics. In grafting experiments, some rat MSCs integrate into the host RPE cell layer of Wistar and RCS rats, indicated by their hexagonal morphology. Subretinally transplanted cells express the epithelial marker cytokeratin and establish tight junctions with the host RPE cells. Furthermore, rescue effects can be demonstrated after grafting of vector-transduced and nontransduced MSCs in semithin sections of dystrophic retinas. Ultrastructurally, MSCs can be detected on top of host RPE and in close contact with photoreceptor outer segments phagocytosing rod outer segments.CONCLUSIONS. Taken together, these results raise the possibility that MSCs have the potency to replace diseased RPE cells and deliver therapeutic proteins into the subretinal space to protect photoreceptor cells from degeneration.