Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
PLoS One. 2013 Aug 19;8(8):e71855. doi: 10.1371/journal.pone.0071855. eCollection 2013.
Degeneration of retinal ganglion cells (RGCs) is a common occurrence in several eye diseases. This study examined the functional improvement and protection of host RGCs in addition to the survival, integration and neuronal differentiation capabilities of anterior specified neural progenitors (NPs) following intravitreal transplantation.
METHODOLOGY/PRINCIPAL FINDINGS: NPs were produced under defined conditions from human induced pluripotent stem cells (hiPSCs) and transplanted into rats whose optic nerves have been crushed (ONC). hiPSCs were induced to differentiate into anterior specified NPs by the use of Noggin and retinoic acid. The hiPSC-NPs were labeled by green fluorescent protein or a fluorescent tracer 1,1' -dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) and injected two days after induction of ONC in hooded rats. Functional analysis according to visual evoked potential recordings showed significant amplitude recovery in animals transplanted with hiPSC-NPs. Retrograde labeling by an intra-collicular DiI injection showed significantly higher numbers of RGCs and spared axons in ONC rats treated with hiPSC-NPs or their conditioned medium (CM). The analysis of CM of hiPSC-NPs showed the secretion of ciliary neurotrophic factor, basic fibroblast growth factor, and insulin-like growth factor. Optic nerve of cell transplanted groups also had increased GAP43 immunoreactivity and myelin staining by FluoroMyelin™ which imply for protection of axons and myelin. At 60 days post-transplantation hiPSC-NPs were integrated into the ganglion cell layer of the retina and expressed neuronal markers.
CONCLUSIONS/SIGNIFICANCE: The transplantation of anterior specified NPs may improve optic nerve injury through neuroprotection and differentiation into neuronal lineages. These NPs possibly provide a promising new therapeutic approach for traumatic optic nerve injuries and loss of RGCs caused by other diseases.
视网膜神经节细胞（RGC）的变性是几种眼病的常见现象。本研究通过玻璃体内移植，研究了前体细胞（NPs）在体内的存活、整合和神经元分化能力以外，宿主 RGC 的功能改善和保护情况。
方法/主要发现：在严格条件下，我们从人诱导多能干细胞（hiPSCs）中产生 NPs，并将其移植到视神经受压（ONC）的大鼠体内。我们使用 Noggin 和维甲酸诱导 hiPSC 分化为前体细胞。hiPSC-NPs 被绿色荧光蛋白或荧光示踪剂 1,1'-二辛基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐（DiI）标记，并在诱导 ONC 后两天注入 Hooded 大鼠。根据视觉诱发电位记录进行的功能分析显示，移植 hiPSC-NPs 的动物的振幅有明显恢复。通过脑室内 DiI 注射进行逆行标记，发现用 hiPSC-NPs 或其条件培养基（CM）处理的 ONC 大鼠的 RGCs 和保留轴突数量显著增加。对 hiPSC-NPs 的 CM 分析表明，睫状神经营养因子、碱性成纤维细胞生长因子和胰岛素样生长因子的分泌增加。细胞移植组的视神经也有更多的 GAP43 免疫反应性和 FluoroMyelin™ 髓鞘染色，这意味着对轴突和髓鞘的保护。移植后 60 天，hiPSC-NPs 整合到视网膜神经节细胞层并表达神经元标记物。
结论/意义：前体细胞的移植可能通过神经保护和向神经元谱系分化来改善视神经损伤。这些 NPs 可能为外伤性视神经损伤和其他疾病引起的 RGC 丧失提供一种有前途的新治疗方法。
