Spatial and Temporal Development of Müller Glial Cells in hiPSC-Derived Retinal Organoids Facilitates the Cell Enrichment and Transcriptome Analysis.
Spatial and Temporal Development of Müller Glial Cells in hiPSC-Derived Retinal Organoids Facilitates the Cell Enrichment and Transcriptome Analysis.
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hiPSC 来源的视网膜类器官中 Müller 胶质细胞的空间和时间发育促进细胞富集和转录组分析。
DOI：
10.3389/fncel.2022.820396
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发表时间：
2022
影响因子：
5.3
通讯作者：
中科院分区：
文献类型：
--
作者：
Müller胶质细胞（MGCs）在视网膜的生理和病理过程中起着重要作用。然而，人MGCs在体内的发育过程尚不清楚，如何获得大量高质量的人MGCs面临技术挑战，阻碍了MGCs的进一步研究和应用。具有所有视网膜细胞亚型的人诱导多能干细胞（hiPSC）衍生的视网膜类器官（RO）为视网膜发育和病症的研究提供了无限的细胞资源和平台。本研究探索了人MGCs在hiPSC衍生的RO中的发育，并开发了一种选择和扩增诱导的MGCs（iMGCs）的方法。在RO中，视网膜祖细胞在分化第（D）60天至D90天期间逐渐分化为S 0X 9 + Ki 67- MGC前体，而表达标记物CRALBP和GS的成熟MGC自D120起逐渐出现，其跨越神经视网膜层的整个厚度。从年龄大于120天的RO中分离的细胞是富集具有高纯度和扩增能力的iMGCs的最佳来源。该细胞在形态、结构、分子和功能等方面均具有人MGCs的典型特征，可连续传代10次以上，在短时间内获得大量细胞。还揭示了扩增的iMGCs的转录组模式。本研究首次阐明了RO模型中人MGC发育的时间过程，其中iMGCs可以富集和扩增，为下游研究和应用于MGC相关视网膜疾病铺平了道路。
Müller glial cells (MGCs) play important roles in human retina during physiological and pathological conditions. However, the development process of human MGCs in vivo remains unclear, and how to obtain large numbers of human MGCs with high quality faces technical challenges, which hinder the further study and application of MGCs. Human induced pluripotent stem cell (hiPSC)-derived retinal organoids (ROs) with all retinal cell subtypes provide an unlimited cell resource and a platform for the studies of retinal development and disorders. This study explored the development of human MGCs in hiPSC-derived ROs and developed an approach to select and expand the induced MGCs (iMGCs). In ROs, retinal progenitor cells progressively differentiated into SOX9+ Ki67– MGC precursors during differentiation day (D) 60 to D90, while mature MGCs expressing markers CRALBP and GS gradually appeared since D120, which spanned the entire thickness of the neural retina layer. Cells isolated from ROs aged older than 120 days was an optimal source for the enrichment of iMGCs with high purity and expansion ability. They had typical features of human MGCs in morphological, structural, molecular and functional aspects, and could be passaged serially at least 10 times, yielding large numbers of cells in a short period. The transcriptome pattern of the expanded iMGCs was also revealed. This study firstly clarified the timecourse of human MGC development in the RO model, where the iMGCs could be enriched and expanded, paving the way for downstream investigation and application in MGC-related retinal disorders.
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影响因子：
25
作者：
Fischer, AJ;Reh, TA
通讯作者：
Reh, TA
DOI：
10.1073/pnas.0807453105
发表时间：
2008-12-09
影响因子：
11.1
作者：
Karl, Mike O.;Hayes, Susan;Reh, Thomas A.
通讯作者：
Reh, Thomas A.
影响因子：
6.2
作者：
Couturier A;Blot G;Vignaud L;Nanteau C;Slembrouck-Brec A;Fradot V;Acar N;Sahel JA;Tadayoni R;Thuret G;Sennlaub F;Roger JE;Goureau O;Guillonneau X;Reichman S
通讯作者：
Reichman S
影响因子：
4.6
作者：
Fligor CM;Langer KB;Sridhar A;Ren Y;Shields PK;Edler MC;Ohlemacher SK;Sluch VM;Zack DJ;Zhang C;Suter DM;Meyer JS
通讯作者：
Meyer JS
DOI：
10.1073/pnas.1901572116
发表时间：
2019-05-28
影响因子：
11.1
作者：
Kim, Sangbae;Lowe, Albert;Liu, Wei
通讯作者：
Liu, Wei