June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Multimodal Photoacoustic Microscopy, OCT, and Fluorescence Imaging for Tracking of Human Induced Pluripotent Stem Cell Derived RPE In Vivo
Author Affiliations & Notes
  • Phuc Nguyen
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Athanasios J Karoukis
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Wei Qian
    Imra America Inc, Ann Arbor, Michigan, United States
  • Lisheng Chen
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • N Dayanthi Perera
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Dongshan Yang
    Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, United States
  • Qitao Zhang
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Josh Zhe
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Jessica Henry
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Bing Liu
    Imra America Inc, Ann Arbor, Michigan, United States
  • Wei Zhang
    Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, United States
  • Abigail T Fahim
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Xueding Wang
    Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, United States
  • Yannis M. Paulus
    Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan, United States
  • Footnotes
    Commercial Relationships   Phuc Nguyen None; Athanasios Karoukis None; Wei Qian None; Lisheng Chen None; N Perera None; Dongshan Yang None; Qitao Zhang None; Josh Zhe None; Jessica Henry None; Bing Liu None; Wei Zhang None; Abigail Fahim None; Xueding Wang None; Yannis Paulus None
  • Footnotes
    Support  National Eye Institute Grant (YMP: 1K08EY027458, 1R01EY033000), Fight for Sight- International Retinal Research Foundation (YMP: FFSGIA16002)
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3828. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Phuc Nguyen, Athanasios J Karoukis, Wei Qian, Lisheng Chen, N Dayanthi Perera, Dongshan Yang, Qitao Zhang, Josh Zhe, Jessica Henry, Bing Liu, Wei Zhang, Abigail T Fahim, Xueding Wang, Yannis M. Paulus; Multimodal Photoacoustic Microscopy, OCT, and Fluorescence Imaging for Tracking of Human Induced Pluripotent Stem Cell Derived RPE In Vivo. Invest. Ophthalmol. Vis. Sci. 2023;64(8):3828.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : We developed a multimodal photoacoustic microscopy (PAM), optical coherence tomography (OCT), and fluorescence microscopy imaging system to visualize the growth, migration, and replacement of damaged RPE after subretinal transplantation.

Methods : Three New Zealand white rabbits received subretinal injection of human induced pluripotent stem cell derived RPE (hiPSC-RPE) cells. The cells were labeled with100 µg/mL ultrapure chain-like gold nanoparticle clusters conjugated with indocyanine green and RGD peptide. RPE damage was performed by laser photocoagulation at a power of 800 mW, beam size of 500 µm, and pulse duration of 0.1 s. The rabbits were treated with twelve spots in each eye and received subretinal injection of 30 µL(3.3×106 cells/ µL) labeled cells. The growth of hiPSC-RPE cells were monitored up to 6 months after transplantation by color photography, PAM, OCT, and fluorescent imaging (FI). All the animal experiments were conducted under a protocol approved by the UM IACUC.

Results : Labeled hiPSC-RPE cells showed normal morphology and function in vitro. The transplanted hiPSC-RPE cells were clearly visualized by fluorescent imaging immediately after the injection and up to 28 days post-injection. The cells gradually localized to the laser injury sites. These cells were clearly observed on OCT images. By obtaining PAM images at 650 nm, the transplanted hiPSC-RPE cells were distinguished from the surrounding microvasculature obtained at 578 nm with strong PA signals due to the strong absorption of hemoglobin. Overlaying PAM and OCT images confirmed the migration of hiPSC-RPE cells to the damaged RPE areas. Histological and immunofluorescent images revealed the presence of grafted hiPSC-RPE cells in the subretinal space with the formation of melanin pigmentation, which is consistent with in vivo imaging.

Conclusions : This study demonstrates a unique technology for longitudinally tracking transplanted cells in living animals using PAM, OCT, and FI imaging. The viability and migration pattern of hiPSC-RPE cells can be monitored and differentiated with contrast enhanced PAM and OCT signal. The labeled cells can be tracked longitudinally over 6 months in the subretinal space. This imaging platform could serve as a potential tool for studies of cell-based therapeutics in different retinal diseases.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

 

Multimodal imaging of hiPSC-RPE cells in living rabbits

Multimodal imaging of hiPSC-RPE cells in living rabbits

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×