Transplantation of cells from eye-like structures differentiated from embryonic stem cells in vitro and in vivo regeneration of retinal ganglion-like cells
Transplantation of cells from eye-like structures differentiated from embryonic stem cells in vitro and in vivo regeneration of retinal ganglion-like cells
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DOI：
10.1007/s00417-007-0710-6
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发表时间：
2008-02-01
影响因子：
2.7
通讯作者：
Kunisada, Takahiro
中科院分区：
文献类型：
--
作者：
Aoki, Hitomi;Hara, Akira;Kunisada, Takahiro
本实验室构建了由神经视网膜谱系细胞、视网膜色素上皮细胞和晶状体细胞组成的胚胎干细胞（ES）衍生的眼样结构。我们已经证明，这些眼状结构的细胞可以整合到小鸡发育中的视神经囊泡中。本研究的目的是确定这些眼状结构的细胞在移植到受伤的成年小鼠视网膜玻璃体后能否分化为视网膜神经节细胞（RGCs）。方法体外诱导胚胎干细胞分化成眼样结构，培养6 d或11 d。移植前向受体小鼠眼内注射NMDA损伤RGCs。假治疗的眼睛接受了相同数量的载体载体。从眼样结构中提取细胞，移植到受损眼和对照眼的玻璃体中。移植后10天或8周采用免疫组化方法对宿主眼进行定性和定量分析。结果胚胎干细胞衍生的眼样结构细胞被整合到RGC层，并在移植到对照（非nmda处理）成人眼时分化为神经元。然而，它们很少表达RGC标记物。当它们被移植到NMDA处理过的眼睛中时，这些细胞在视网膜表面扩散，覆盖了被NMDA损伤的宿主RGC层的相对大面积。胚胎干细胞来源的眼细胞经常分化为表达RGC特异性标记的细胞，并形成新的RGC层。此外，观察到少量这些ES细胞衍生的细胞向宿主视盘延伸轴突样突起。然而，视觉诱发的反应不能从视觉皮层记录下来。这些发现表明，胚胎干细胞衍生的眼样结构包含可以分化为rg样细胞并再生新的RGC层的细胞。这些细胞似乎也融入视网膜，并向视神经头延伸轴突样突。
Bcakground An embryonic stem (ES) cell-derived eye-like structure, made up of neural retinal lineage cells, retinal pigment epithelial (RPE) cells, and lens cells was constructed in our laboratory. We have shown that cells from these eye-like structures can be integrated into the developing optic vesicle of chicks. The purpose of this study was to determine whether the cells from these eye-like structures can differentiate into retinal ganglion cells (RGCs) when transplanted into the vitreous of an injured adult mouse retina.Methods ES cells were induced to differentiate into eye-like structures in vitro for 6 or 11 days. Recipient mouse eyes were injected with NMDA to injure the RGCs prior to the transplantation. Sham-treated eyes received the same amount of carrier vehicle. Cells were extracted from the eye-like structures and transplanted into the vitreous of damaged and control eyes. The host eyes were analyzed both qualitatively and quantitatively by immunohistochemistry 10 days or 8 weeks after transplantation.Results Cells from the ES cell-derived eye-like structures were integrated into the RGC layer, and differentiated into neurons when transplanted into control (non-NMDA-treated) adult eyes. However, they rarely expressed RGC markers. When they were transplanted into NMDA-treated eyes, the cells spread on the surface of the retina and covered a relatively large area of the host RGC layer that had been injured by the NMDA. The cells from the ES cell-derived eye cells frequently differentiated into cells expressing RGC-specific markers, and formed a new RGC layer. In addition, a small number of these ES cell-derived cells were observed to extend axon-like processes toward the optic disc of the host. However, visually evoked responses could not be recorded from the visual cortex.DiscussionThese findings suggest that ES cell-derived eye-like structures contain cells that can differentiate into RG-like cells and regenerate a new RGC layer. These cells also appeared to be integrated into the retina and extend axon-like processes toward the optic nerve head.