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Single-cell transcriptomic analysis of corneal organoids during development.

Aditi Swarup1, Ragini Phansalkar1, Maurizio Morri2

  • 1Department of Ophthalmology, Stanford University School of Medicine, Stanford, CA, USA.

Stem Cell Reports
|December 1, 2023
PubMed
Summary

Human induced pluripotent stem cell-derived corneal organoids show distinct cellular development over four months. Four-month-old organoids closely mimic the cellular complexity of fetal and adult corneas, offering disease modeling potential.

Keywords:
Corneal organoidsdevelopmentsingle cell RNAseq

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Area of Science:

  • Ophthalmology
  • Developmental Biology
  • Stem Cell Biology

Background:

  • Corneal organoids are valuable for disease modeling and transplantation.
  • Their developmental maturation process remains incompletely understood.

Purpose of the Study:

  • To characterize the cellular heterogeneity of human induced pluripotent stem cell (iPSC)-derived corneal organoids during maturation.
  • To compare organoid development with fetal corneal development.

Main Methods:

  • Single-cell RNA sequencing was employed to analyze corneal organoids at 1, 2, 3, and 4 months of development.
  • RNA velocity trajectory analysis was used to infer cell differentiation pathways.

Main Results:

  • Pluripotent cell clusters committed to the epithelial lineage were observed at 1 month.
  • Early markers for corneal epithelial, endothelial, and stromal cells appeared at 2 months.
  • Keratocytes became the predominant cell type by 3 months, followed by a dominant epithelial cell population at 4 months.
  • Four-month-old organoids exhibited cellular complexity comparable to fetal (16 weeks post-conception) and adult corneas.
  • RNA velocity analysis indicated that less differentiated cells give rise to corneal epithelial cells.

Conclusions:

  • Human iPSC-derived corneal organoids recapitulate key stages of corneal development.
  • Mature corneal organoids serve as a robust model for studying corneal biology and disease.
  • This study provides insights into the differentiation dynamics of corneal cell types within organoids.