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Episodic live imaging of cone photoreceptor maturation in GNAT2-EGFP retinal organoids.
Bai, Jinlun; Koos, David S; Stepanian, Kayla; Fouladian, Zachary; Shayler, Dominic W H; Aparicio, Jennifer G; Fraser, Scott E; Moats, Rex A; Cobrinik, David.
Afiliación
  • Bai J; The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Koos DS; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Stepanian K; Development, Stem Cell, and Regenerative Medicine Program, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
  • Fouladian Z; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Shayler DWH; Translational Biomedical Imaging Laboratory, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Aparicio JG; Department of Radiology, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Fraser SE; The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Moats RA; The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA, USA.
  • Cobrinik D; The Vision Center, Department of Surgery, Children's Hospital Los Angeles, Los Angeles, CA, USA.
bioRxiv ; 2023 Mar 01.
Article en En | MEDLINE | ID: mdl-36909527
ABSTRACT
Fluorescent reporter pluripotent stem cell (PSC) derived retinal organoids are powerful tools to investigate cell type-specific development and disease phenotypes. When combined with live imaging, they enable direct and repeated observation of cell behaviors within a developing retinal tissue. Here, we generated a human cone photoreceptor reporter line by CRISPR/Cas9 genome editing of WTC11-mTagRFPT-LMNB1 human induced pluripotent stem cells (iPSCs) by inserting enhanced green fluorescent protein (EGFP) coding sequences and a 2A self-cleaving peptide at the N-terminus of Guanine Nucleotide-Binding Protein Subunit Alpha Transducin 2 (GNAT2). In retinal organoids generated from these iPSCs, the GNAT2-EGFP allele robustly and exclusively labeled both immature and mature cones starting at culture day 34. Episodic confocal live imaging of hydrogel immobilized retinal organoids allowed tracking of morphological maturation of individual cones for >18 weeks and revealed inner segment accumulation of mitochondria and growth at 12.2 cubic microns per day from day 126 to day 153. Immobilized GNAT2-EGFP cone reporter organoids provide a valuable tool for investigating human cone development and disease.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Idioma: En Revista: BioRxiv Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Idioma: En Revista: BioRxiv Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos