Human-Induced Pluripotent Stem Cells Generate Light Responsive Retinal Organoids with Variable and Nutrient-Dependent Efficiency.
Human-Induced Pluripotent Stem Cells Generate Light Responsive Retinal Organoids with Variable and Nutrient-Dependent Efficiency.
复制标题
人类诱导的多能干细胞产生具有可变和养分依赖性效率的光响应性视网膜器官。
DOI：
10.1002/stem.2883
复制
发表时间：
2018-10
期刊：
影响因子：
--
通讯作者：
Lako M
中科院分区：
文献类型：
--
作者：
Hallam D;Hilgen G;Dorgau B;Zhu L;Yu M;Bojic S;Hewitt P;Schmitt M;Uteng M;Kustermann S;Steel D;Nicholds M;Thomas R;Treumann A;Porter A;Sernagor E;Armstrong L;Lako M
用于进行药理学和毒理学研究的人视网膜体外模型的可用性是一个迫切且未满足的需求。开发人类视网膜体外模型的一个重要步骤是能够从可再生和患者特异性来源产生层压的、生理功能性的和光响应性视网膜类器官。我们研究了五种不同的人诱导多能干细胞（iPSC）系，并显示其产生视网膜类器官的效率存在显著差异。尽管存在这种变异性，但到分化的第5个月，所有iPSC衍生的视网膜类器官都能够产生光反应，尽管不成熟，但与新生小鼠视网膜记录的最早光反应相当，接近睁眼期。此时，所有iPSC衍生的视网膜类器官均表现出形态良好的外核样层，其中包含具有内节、连接纤毛和外核样节的光感受器。分化过程高度依赖于种子细胞密度和养分的可利用性由析因实验设计确定。我们采用了多孔板形式的分化方案，其增强了具有视网膜色素上皮（RPE）的视网膜类器官的产生，并改善了神经节细胞发育和对生理刺激的反应。我们测试了iPSC衍生的视网膜类器官对莫西沙星的反应，并表明与体内成年小鼠视网膜相似，主要受影响的细胞类型是光感受器。总之，我们的数据表明，衍生自精心选择和分化有效的iPSC系的光响应性视网膜类器官可以以药理学和药物筛选目的所需的规模产生。干细胞2018;36：1535-1551
The availability of in vitro models of the human retina in which to perform pharmacological and toxicological studies is an urgent and unmet need. An essential step for developing in vitro models of human retina is the ability to generate laminated, physiologically functional, and light‐responsive retinal organoids from renewable and patient specific sources. We investigated five different human‐induced pluripotent stem cell (iPSC) lines and showed a significant variability in their efficiency to generate retinal organoids. Despite this variability, by month 5 of differentiation, all iPSC‐derived retinal organoids were able to generate light responses, albeit immature, comparable to the earliest light responses recorded from the neonatal mouse retina, close to the period of eye opening. All iPSC‐derived retinal organoids exhibited at this time a well‐formed outer nuclear like layer containing photoreceptors with inner segments, connecting cilium, and outer like segments. The differentiation process was highly dependent on seeding cell density and nutrient availability determined by factorial experimental design. We adopted the differentiation protocol to a multiwell plate format, which enhanced generation of retinal organoids with retinal‐pigmented epithelium (RPE) and improved ganglion cell development and the response to physiological stimuli. We tested the response of iPSC‐derived retinal organoids to Moxifloxacin and showed that similarly to in vivo adult mouse retina, the primary affected cell types were photoreceptors. Together our data indicate that light responsive retinal organoids derived from carefully selected and differentiation efficient iPSC lines can be generated at the scale needed for pharmacology and drug screening purposes. stem cells 2018;36:1535–1551
登录
查看更多内容
DOI：
10.1126/science.aao3130
发表时间：
2017-10-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Drost J;van Boxtel R;Blokzijl F;Mizutani T;Sasaki N;Sasselli V;de Ligt J;Behjati S;Grolleman JE;van Wezel T;Nik-Zainal S;Kuiper RP;Cuppen E;Clevers H
通讯作者：
Clevers H
影响因子：
16.6
作者：
Arno, Benedetta;Grassivaro, Francesca;Muzio, Luca
通讯作者：
Muzio, Luca
影响因子：
--
作者：
Holman, Jerry D;Tabb, David L;Mallick, Parag
通讯作者：
Mallick, Parag
影响因子：
64.5
作者：
Huch M;Gehart H;van Boxtel R;Hamer K;Blokzijl F;Verstegen MM;Ellis E;van Wenum M;Fuchs SA;de Ligt J;van de Wetering M;Sasaki N;Boers SJ;Kemperman H;de Jonge J;Ijzermans JN;Nieuwenhuis EE;Hoekstra R;Strom S;Vries RR;van der Laan LJ;Cuppen E;Clevers H
通讯作者：
Clevers H
影响因子：
5.9
作者：
Gonzalez-Cordero A;Kruczek K;Naeem A;Fernando M;Kloc M;Ribeiro J;Goh D;Duran Y;Blackford SJI;Abelleira-Hervas L;Sampson RD;Shum IO;Branch MJ;Gardner PJ;Sowden JC;Bainbridge JWB;Smith AJ;West EL;Pearson RA;Ali RR
通讯作者：
Ali RR