Generation of Retinal Ganglion Cells With Functional Axons From Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells.
Generation of Retinal Ganglion Cells With Functional Axons From Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells.
复制标题
DOI：
10.1167/iovs.16-19166
复制
发表时间：
2016-06
影响因子：
4.4
通讯作者：
Taku Tanaka;T. Yokoi;F. Tamalu;Shu-ichi Watanabe;S. Nishina;N. Azuma
中科院分区：
文献类型：
--
作者：
Taku Tanaka;T. Yokoi;F. Tamalu;Shu-ichi Watanabe;S. Nishina;N. Azuma
我们先前从人诱导多能干细胞（hiPSC）中产生了具有功能轴突的自诱导视网膜神经节细胞（RGC）。我们研究了来自小鼠胚胎干细胞（mESC）和诱导多能干细胞（miPSC）的自诱导RGC是否通过类似的诱导方案实现。方法采用三维培养法诱导视网膜神经节细胞分化为视网膜细胞系，并在二维培养皿中加入脑源性神经营养因子（BDNF）。结果视网膜神经节细胞生长于视泡周围的细胞团中，轴突多从视泡周围的RGC胞体中生长出来。RGCs的分化通过特异性标志物（包括Brn3a和Math5）的表达来证实。轴突含有神经丝亚型和tau蛋白，并表现出轴突运输和钠依赖性动作电位。来源于mESC和miPSC的RGC通常显示出相似的特征，包括RNA和蛋白质表达水平和功能。结论mESC和miPSC，特别是后者，产生的视网膜神经节细胞可能有助于与RGCs相关的研究和转基因小鼠的体外分析。
PURPOSE We previously generated self-induced retinal ganglion cells (RGCs) with functional axons from human induced pluripotent stem cells (hiPSCs). We investigated whether self-induced RGCs from mouse embryonic stem cells (mESCs) and induced pluripotent stem cells (miPSCs) are realized by the similar induction protocol. METHODS Retinal ganglion cells were induced using a protocol in which floating embryoid bodies (EBs) were differentiated into a retinal cell lineage in three-dimensional culture and subsequently attached to two-dimensional culture dishes with brain-derived neurotrophic factor (BDNF) supplementation. RESULTS Retinal ganglion cells developed in an attached clump of cells originating from the optic vesicle, and most axons grew from RGC cell bodies at the margins of the clump. The differentiation of RGCs was confirmed by the expression of specific markers, including Brn3a and Math5. The axons contained neurofilament subtypes and tau, and manifested axonal transport and sodium-dependent action potentials. The RGCs derived from mESCs and miPSCs generally showed similar profiles, including RNA and protein expression levels and function. CONCLUSIONS Retinal ganglion cells generated from mESCs and miPSCs, especially the latter, may contribute to research associated with RGCs and to in vitro analyses of genetically modified mice.