Retinal Organoid Microenvironment Enhanced Bioactivities of Microglia-Like Cells Derived From HiPSCs.

IF 5 2区 医学 Q1 OPHTHALMOLOGY
Mei-Ling Gao, Tong-Yu Wang, Xin Lin, Chun Tang, Mengyao Li, Zhan-Pei Bai, Zhi-Cong Liu, Li-Jun Chen, Qing-Ran Kong, Shao-Hui Pan, Shan-Shan Zeng, Ya Guo, Jian-Qi Cai, Xiu-Feng Huang, Jun Zhang
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Abstract

Purpose: Microglia-like cells derived from stem cells (iMG) provide a plentiful cell source for studying the functions of microglia in both normal and pathological conditions. Our goal is to establish a simplified and effective method for generating iMG in a precisely defined system. Additionally, we aim to achieve functional maturation of iMG through coculture with retinal organoids.

Methods: In this study, iMG were produced under precisely defined conditions. They were subjected to LPS and poly IC stimulation. Additionally, we examined distinct phenotypic and functional variances between iMG and HMC3, a commonly used human microglia cell line. To investigate how the retinal cell interaction enhances microglial properties, iMG were cocultured with retinal organoids, producing CC-iMG. We performed RNA sequencing, electrophysiological analysis, and transmission electron microscope (TEM) to examine the maturation of CC-iMG compared to iMG.

Results: Our results demonstrated that iMG performed immune-responsive profiles closely resembling those of primary human microglia. Compared to HMC3, iMG expressed a higher level of typical microglial markers and exhibited enhanced phagocytic activity. The transcriptomic analysis uncovered notable alterations in the ion channel profile of CC-iMG compared to iMG. Electrophysiological examination demonstrated a heightened intensity of inward- and outward-rectifying K+ currents in CC-iMG. Furthermore, CC-iMG displayed elevated numbers of lysosomes and mitochondria, coupled with increased phagocytic activity.

Conclusions: These findings contribute to advancing our understanding of human microglial biology, specifically in characterizing and elucidating the functions of CC-iMG, thereby offering an in vitro microglial model for future scientific research and potential clinical applications in cell therapy.

视网膜类器官微环境增强了 HiPSCs 衍生的小胶质细胞的生物活性。
目的:源自干细胞的小胶质细胞样细胞(iMG)为研究小胶质细胞在正常和病理情况下的功能提供了丰富的细胞来源。我们的目标是建立一种简化而有效的方法,在精确定义的系统中生成 iMG。此外,我们还希望通过与视网膜器官组织共培养实现 iMG 的功能成熟:在这项研究中,iMG 是在精确定义的条件下产生的。方法:在本研究中,iMG 是在精确定义的条件下产生的,它们受到 LPS 和聚 IC 的刺激。此外,我们还研究了 iMG 和 HMC3(一种常用的人类小胶质细胞系)之间不同的表型和功能差异。为了研究视网膜细胞相互作用如何增强小胶质细胞的特性,我们将 iMG 与视网膜器官组织共培养,产生了 CC-iMG。我们进行了 RNA 测序、电生理分析和透射电子显微镜(TEM),以研究 CC-iMG 与 iMG 相比的成熟过程:结果:我们的研究结果表明,iMG的免疫反应谱与原代人类小胶质细胞的免疫反应谱非常相似。与 HMC3 相比,iMG 表达了更高水平的典型小胶质细胞标记物,并表现出更强的吞噬活性。转录组分析发现,与 iMG 相比,CC-iMG 的离子通道特征发生了显著变化。电生理学检查显示,CC-iMG 的内向和外向校正 K+ 电流强度增加。此外,CC-iMG 还显示溶酶体和线粒体数量增加,同时吞噬活性增强:这些发现有助于增进我们对人类小胶质细胞生物学的了解,特别是在表征和阐明 CC-iMG 的功能方面,从而为未来的科学研究和细胞疗法的潜在临床应用提供了一种体外小胶质细胞模型。
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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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