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

  • Invest Ophthalmol Vis Sci. 2024 Oct 1;65(12):19. doi: 10.1167/iovs.65.12.19.
Mei-Ling Gao  1  2 ,  Tong-Yu Wang  1  2 ,  Xin Lin  1  2 ,  Chun Tang  1  2 ,  Mengyao Li  3 ,  Zhan-Pei Bai  4 ,  Zhi-Cong Liu  1  2 ,  Li-Jun Chen  5 ,  Qing-Ran Kong  5 ,  Shao-Hui Pan  1 ,  Shan-Shan Zeng  6 ,  Ya Guo  6 ,  Jian-Qi Cai  6 ,  Xiu-Feng Huang  4 ,  Jun Zhang  1  2
Affiliations
  • 1. The State Key Laboratory of Ophthalmology, Optometry and Visual Science, Wenzhou Medical University, Wenzhou, China.
  • 2. Laboratory of Retinal Physiology and Disease, Eye Hospital and School of Ophthalmology and Optometry, Wenzhou Medical University, Wenzhou, China.
  • 3. Department of Critical Care Medicine, The Second Clinical Medical College, Jinan University (Shenzhen People's Hospital), Shenzhen, China.
  • 4. Zhejiang Provincial Clinical Research Center for Pediatric Disease, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
  • 5. School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China.
  • 6. China National Institute of Standardization, Beijing, China.
PMID: 39392440 DOI: 10.1167/iovs.65.12.19
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.

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