Department of Ophthalmology, Columbia University, New York City, NY, USA.
Transplantation. 2010 Apr 27;89(8):911-9. doi: 10.1097/TP.0b013e3181d45a61.
To study whether C57BL/6J-Tyr/J (C2J) mouse embryonic stem (ES) cells can differentiate into retinal pigment epithelial (RPE) cells in vitro and then restore retinal function in a model for retinitis pigmentosa: Rpe65/Rpe65 C57BL6 mice.
Yellow fluorescent protein (YFP)-labeled C2J ES cells were induced to differentiate into RPE-like structures on PA6 feeders. RPE-specific markers are expressed from differentiated cells in vitro. After differentiation, ES cell-derived RPE-like cells were transplanted into the subretinal space of postnatal day 5 Rpe65/Rpe65 mice. Live imaging of YFP-labeled C2J ES cells demonstrated survival of the graft. Electroretinograms (ERGs) were performed on transplanted mice to evaluate the functional outcome of transplantation.
RPE-like cells derived from ES cells sequentially express multiple RPE-specific markers. After transplantation, YFP-labeled cells can be tracked with live imaging for as long as 7 months. Although more than half of the mice were complicated with retinal detachments or tumor development, one fourth of the mice showed increased electroretinogram responses in the transplanted eyes. Rpe65/Rpe65 mice transplanted with RPE-like cells showed significant visual recovery during a 7-month period, whereas those injected with saline, PA6 feeders, or undifferentiated ES cells showed no rescue.
ES cells can differentiate, morphologically, and functionally, into RPE-like cells. Based on these findings, differentiated ES cells have the potential for the development of new therapeutic approaches for RPE-specific diseases such as certain forms of retinitis pigmentosa and macular degeneration. Nevertheless, stringent control of retinal detachment and teratoma development will be necessary before initiation of treatment trials.
研究 C57BL/6J-Tyr/J（C2J）小鼠胚胎干细胞（ES 细胞）是否能在体外分化为视网膜色素上皮（RPE）细胞，然后在 Rpe65/Rpe65 C57BL6 小鼠的模型中恢复视网膜功能：色素性视网膜炎。
用黄色荧光蛋白（YFP）标记的 C2J ES 细胞在 PA6 饲养层上诱导分化为 RPE 样结构。体外分化的细胞表达 RPE 特异性标记物。分化后，将 ES 细胞来源的 RPE 样细胞移植到出生后第 5 天 Rpe65/Rpe65 小鼠的视网膜下腔。用活细胞成像技术观察 YFP 标记的 C2J ES 细胞的移植细胞存活情况。对移植小鼠进行视网膜电图（ERG）检查，以评估移植的功能结果。
ES 细胞分化而来的 RPE 样细胞依次表达多种 RPE 特异性标记物。移植后，YFP 标记的细胞可以通过活体成像追踪长达 7 个月。尽管超过一半的小鼠伴有视网膜脱离或肿瘤发生，但有四分之一的小鼠在移植眼中的视网膜电图反应增加。移植 RPE 样细胞的 Rpe65/Rpe65 小鼠在 7 个月的时间内显示出明显的视力恢复，而注射生理盐水、PA6 饲养层或未分化的 ES 细胞的小鼠则没有恢复。
ES 细胞可以在形态和功能上分化为 RPE 样细胞。基于这些发现，分化的 ES 细胞有可能为特定形式的色素性视网膜炎和黄斑变性等 RPE 特异性疾病开发新的治疗方法。然而，在开始治疗试验之前，必须严格控制视网膜脱离和畸胎瘤的发生。
