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Structural and Functional Characterization of Human Stem-Cell-Derived Retinal Organoids by Live Imaging.
Browne, Andrew W; Arnesano, Cosimo; Harutyunyan, Narine; Khuu, Thien; Martinez, Juan Carlos; Pollack, Harvey A; Koos, David S; Lee, Thomas C; Fraser, Scott E; Moats, Rex A; Aparicio, Jennifer G; Cobrinik, David.
Afiliação
  • Browne AW; USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, California, United States.
  • Arnesano C; Translational Imaging Center, University of Southern California, Los Angeles, California, United States 3Department of Molecular and Computational Biology, University of Southern California, Los Angeles, California, United States.
  • Harutyunyan N; The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Khuu T; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Martinez JC; USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, California, United States.
  • Pollack HA; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California, United States 6Department of Radiology, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Koos DS; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California, United States 6Department of Radiology, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Lee TC; USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, California, United States 4The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Fraser SE; Translational Imaging Center, University of Southern California, Los Angeles, California, United States 3Department of Molecular and Computational Biology, University of Southern California, Los Angeles, California, United States 5The Saban Research Institute, Children's Hospital Los Angeles, Los An
  • Moats RA; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California, United States 6Department of Radiology, Children's Hospital Los Angeles, Los Angeles, California, United States 7Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern Califor
  • Aparicio JG; The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, California, United States.
  • Cobrinik D; USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, California, United States 4The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, California, United States 5The Saban Research Instit
Invest Ophthalmol Vis Sci ; 58(9): 3311-3318, 2017 07 01.
Article em En | MEDLINE | ID: mdl-28672397
ABSTRACT

Purpose:

Human pluripotent stem cell (hPSC)-derived retinal organoids are a platform for investigating retinal development, pathophysiology, and cellular therapies. In contrast to histologic analysis in which multiple specimens fixed at different times are used to reconstruct developmental processes, repeated analysis of the same living organoids provides a more direct means to characterize changes. New live imaging modalities can provide insights into retinal organoid structure and metabolic function during in vitro growth. This study employed live tissue imaging to characterize retinal organoid development, including metabolic changes accompanying photoreceptor differentiation.

Methods:

Live hPSC-derived retinal organoids at different developmental stages were examined for microanatomic organization and metabolic function by phase contrast microscopy, optical coherence tomography (OCT), fluorescence lifetime imaging microscopy (FLIM), and hyperspectral imaging (HSpec). Features were compared to those revealed by histologic staining, immunostaining, and microcomputed tomography (micro-CT) of fixed organoid tissue.

Results:

We used FLIM and HSpec to detect changes in metabolic activity as organoids differentiated into organized lamellae. FLIM detected increased glycolytic activity and HSpec detected retinol and retinoic acid accumulation in the organoid outer layer, coinciding with photoreceptor genesis. OCT enabled imaging of lamellae formed during organoid maturation. Micro-CT revealed three-dimensional structure, but failed to detect lamellae.

Conclusions:

Live imaging modalities facilitate real-time and nondestructive imaging of retinal organoids as they organize into lamellar structures. FLIM and HSpec enable rapid detection of lamellar structure and photoreceptor metabolism. Live imaging techniques may aid in the continuous evaluation of retinal organoid development in diverse experimental and cell therapy settings.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Retina / Organoides / Células-Tronco Pluripotentes / Técnicas de Diagnóstico Oftalmológico Idioma: En Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Retina / Organoides / Células-Tronco Pluripotentes / Técnicas de Diagnóstico Oftalmológico Idioma: En Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos