3D culture of human pluripotent stem cells in RGD-alginate hydrogel improves retinal tissue development
3D culture of human pluripotent stem cells in RGD-alginate hydrogel improves retinal tissue development
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DOI：
10.1016/j.actbio.2016.11.016
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发表时间：
2017-02-01
影响因子：
9.7
通讯作者：
Lako, Majlinda
中科院分区：
文献类型：
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作者：
Hunt, Nicola C.;Hallam, Dean;Lako, Majlinda
目前还没有有效治疗许多视网膜疾病的方法。视网膜色素上皮（RPE）和神经视网膜可以由人胚胎干细胞/诱导多能干细胞（hESCs/hiPSCs）生成。然而，目前的协议的效力是有限的。假设水凝胶中的三维（3D）培养可以增强hiPSCs/hESCs层压神经视网膜和/或RPE的生成。将hiPSC和hesc衍生的胚状体（EBs）包裹在0.5% rgd -海藻酸盐中；RGD-alginate 1%;透明质酸（HA）或透明质酸/明胶水凝胶维持至第45天。与对照组（无凝胶）相比，0.5% rgd -海藻酸盐增加了：有色素RPE灶的EBs百分比；同时存在视神经囊泡（OVs）和色素RPE的EBs百分比；RPE覆盖的区域；RPE细胞频率（CRALBP-F）；RPE标记物（TYR和RPE65）和视网膜神经节细胞标记物（MATHS）的表达。此外，0.5% rgd -海藻酸盐水凝胶包封对其他神经视网膜标志物（PROX1、CRX、RCVRN、AP2 α或VSX2）的表达（通过qRT-PCR检测）和VSX2阳性细胞的百分比（通过流式细胞术检测）没有不利影响。1% rgd -海藻酸盐增加了伴有OVs和/或RPE的EBs的百分比，但对视网膜分化的任何其他指标没有显著影响。ha基水凝胶对视网膜组织发育无显著影响。结果表明，0.5% rgd -海藻酸盐水凝胶培养可促进hESCs/hiPSCs的视网膜组织分化。这种rgd -海藻酸盐支架可用于神经视网膜和RPE的衍生、运输和移植，也可促进其他色素组织、神经组织或上皮组织的形成。随着世界人口年龄的增长，视网膜疾病的负担也在不断增加。人类多能干细胞（PSCs）视网膜组织移植被认为是一种很有前途的治疗方法。然而，使用已定义的培养基从PSCs中衍生视网膜组织是一个漫长的过程，并且在不同的细胞系之间经常发生变化。该研究表明海藻酸盐水凝胶促进了PSCs的视网膜组织发育，而透明质酸水凝胶则没有。这是第一个表明生物材料支架的3D培养可以改善PSCs的视网膜组织衍生的研究。这些发现表明海藻酸盐水凝胶在视网膜组织的衍生和后续移植中的临床应用潜力。这项工作也可能对其他色素、神经或上皮组织的衍生有启示。Copyright (C) 2016 Elsevier Ltd代表Acta Materialia Inc出版
No treatments exist to effectively treat many retinal diseases. Retinal pigmented epithelium (RPE) and neural retina can be generated from human embryonic stem cells/induced pluripotent stem cells (hESCs/hiPSCs). The efficacy of current protocols is, however, limited. It was hypothesised that generation of laminated neural retina and/or RPE from hiPSCs/hESCs could be enhanced by three dimensional (3D) culture in hydrogels. hiPSC- and hESC-derived embryoid bodies (EBs) were encapsulated in 0.5% RGD-alginate; 1% RGD-alginate; hyaluronic acid (HA) or HA/gelatin hydrogels and maintained until day 45. Compared with controls (no gel), 0.5% RGD-alginate increased: the percentage of EBs with pigmented RPE foci; the percentage EBs with optic vesicles (OVs) and pigmented RPE simultaneously; the area covered by RPE; frequency of RPE cells (CRALBP-F); expression of RPE markers (TYR and RPE65) and the retinal ganglion cell marker, MATHS. Furthermore, 0.5% RGD-alginate hydrogel encapsulation did not adversely affect the expression of other neural retina markers (PROX1, CRX, RCVRN, AP2 alpha or VSX2) as determined by qRT-PCR, or the percentage of VSX2 positive cells as determined by flow cytometry. 1% RGD-alginate increased the percentage of EBs with OVs and/or RPE, but did not significantly influence any other measures of retinal differentiation. HA-based hydrogels had no significant effect on retinal tissue development. The results indicated that derivation of retinal tissue from hESCs/hiPSCs can be enhanced by culture in 0.5% RGD-alginate hydrogel. This RGD-alginate scaffold may be useful for derivation, transport and transplantation of neural retina and RPE, and may also enhance formation of other pigmented, neural or epithelial tissue.Statement of SignificanceThe burden of retinal disease is ever growing with the increasing age of the world-wide population. Transplantation of retinal tissue derived from human pluripotent stem cells (PSCs) is considered a promising treatment. However, derivation of retinal tissue from PSCs using defined media is a lengthy process and often variable between different cell lines.This study indicated that alginate hydrogels enhanced retinal tissue development from PSCs, whereas hyaluronic acid-based hydrogels did not. This is the first study to show that 3D culture with a biomaterial scaffold can improve retinal tissue derivation from PSCs.These findings indicate potential for the clinical application of alginate hydrogels for the derivation and subsequent transplantation retinal tissue. This work may also have implications for the derivation of other pigmented, neural or epithelial tissue. Crown Copyright (C) 2016 Published by Elsevier Ltd on behalf of Acta Materialia Inc.