Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines

Elizabeth E. Capowski, Kayvan Samimi, Steven J. Mayerl, M. Joseph Phillips, Isabel Pinilla, Sara E. Howden, Jishnu Saha, Alex D. Jansen, Kimberly L. Edwards, Lindsey D. Jager, Katherine Barlow, Rasa Valiauga, Zachary Erlichman, Anna Hagstrom, Divya Sinha, Valentin M. Sluch, Xitiz Chamling, Donald J Zack, Melissa C. Skala, David M. Gamm

Research output: Contribution to journalArticle

Abstract

Numerous protocols have been described for producing neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture of 3D organoids. Although nearly all such methods yield at least partial segments of retinal structure with a mature appearance, variabilities exist within and between organoids that can change over a protracted time course of differentiation. Adding to this complexity are potential differences in the composition and configuration of retinal organoids when viewed across multiple differentiations and hPSC lines. In an effort to understand better the current capabilities and limitations of these cultures, we generated retinal organoids from 16 hPSC lines and monitored their appearance and structural organization over time by light microscopy, immunocytochemistry, metabolic imaging and electron microscopy. We also employed optical coherence tomography and 3D imaging techniques to assess and compare whole or broad regions of organoids to avoid selection bias. Results from this study led to the development of a practical staging system to reduce inconsistencies in retinal organoid cultures and increase rigor when utilizing them in developmental studies, disease modeling and transplantation.

Original languageEnglish (US)
JournalDevelopment (Cambridge, England)
Volume146
Issue number1
DOIs
StatePublished - Jan 9 2019

Fingerprint

Organoids
Pluripotent Stem Cells
Cell Line
Selection Bias
Optical Coherence Tomography
Retina
Microscopy
Electron Microscopy
Transplantation
Immunohistochemistry
Light

Keywords

  • Cell culture
  • Differentiation
  • Human pluripotent stem cells
  • Organoids
  • Photoreceptors
  • Retina

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology

Cite this

Capowski, E. E., Samimi, K., Mayerl, S. J., Phillips, M. J., Pinilla, I., Howden, S. E., ... Gamm, D. M. (2019). Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines. Development (Cambridge, England), 146(1). https://doi.org/10.1242/dev.171686

Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines. / Capowski, Elizabeth E.; Samimi, Kayvan; Mayerl, Steven J.; Phillips, M. Joseph; Pinilla, Isabel; Howden, Sara E.; Saha, Jishnu; Jansen, Alex D.; Edwards, Kimberly L.; Jager, Lindsey D.; Barlow, Katherine; Valiauga, Rasa; Erlichman, Zachary; Hagstrom, Anna; Sinha, Divya; Sluch, Valentin M.; Chamling, Xitiz; Zack, Donald J; Skala, Melissa C.; Gamm, David M.

In: Development (Cambridge, England), Vol. 146, No. 1, 09.01.2019.

Research output: Contribution to journalArticle

Capowski, EE, Samimi, K, Mayerl, SJ, Phillips, MJ, Pinilla, I, Howden, SE, Saha, J, Jansen, AD, Edwards, KL, Jager, LD, Barlow, K, Valiauga, R, Erlichman, Z, Hagstrom, A, Sinha, D, Sluch, VM, Chamling, X, Zack, DJ, Skala, MC & Gamm, DM 2019, 'Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines', Development (Cambridge, England), vol. 146, no. 1. https://doi.org/10.1242/dev.171686
Capowski, Elizabeth E. ; Samimi, Kayvan ; Mayerl, Steven J. ; Phillips, M. Joseph ; Pinilla, Isabel ; Howden, Sara E. ; Saha, Jishnu ; Jansen, Alex D. ; Edwards, Kimberly L. ; Jager, Lindsey D. ; Barlow, Katherine ; Valiauga, Rasa ; Erlichman, Zachary ; Hagstrom, Anna ; Sinha, Divya ; Sluch, Valentin M. ; Chamling, Xitiz ; Zack, Donald J ; Skala, Melissa C. ; Gamm, David M. / Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines. In: Development (Cambridge, England). 2019 ; Vol. 146, No. 1.
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