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Retinoid uptake, processing, and secretion in human iPS-RPE support the visual cycle.

Alberto Muñiz1, Whitney A Greene, Mark L Plamper

  • 1Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston, Houston, Texas.

Investigative Ophthalmology & Visual Science
|November 21, 2013
PubMed
Summary
This summary is machine-generated.

Human induced pluripotent stem (iPS) cell-derived retinal pigmented epithelium (iPS-RPE) expresses key visual cycle proteins and demonstrates functional retinoid processing. This confirms iPS-RPE

Keywords:
induced-pluripotent stem cellretinal pigment epitheliumvisual cycle

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Area of Science:

  • Stem cell biology
  • Ophthalmology
  • Biochemistry

Background:

  • Human induced pluripotent stem (iPS) cells offer a potential source for retinal pigmented epithelium (RPE) transplantation.
  • Assessing the functional capacity of iPS-RPE, particularly its role in the visual cycle, is crucial for its therapeutic application.

Purpose of the Study:

  • To investigate the expression of visual cycle proteins in iPS-RPE.
  • To evaluate the functional retinoid processing ability of iPS-RPE.

Main Methods:

  • iPS-RPE derived from human iPS cells.
  • Immunocytochemistry, RT-PCR, and Western blot used to detect visual cycle genes (LRAT, RPE65, CRALBP, PEDF).
  • Functional assessment involved delivering all-trans retinol to cultured cells and homogenates.

Main Results:

  • iPS-RPE expressed key visual cycle genes: LRAT, CRALBP, and RPE65.
  • Synthesized all-trans retinyl esters upon all-trans retinol incubation, with synthesis inhibited by NEM, confirming LRAT activity.
  • Released 11-cis retinaldehyde into culture media, indicating functional retinoid processing.

Conclusions:

  • iPS-RPE exhibits RPE characteristics and expresses essential visual cycle proteins.
  • The cells possess the necessary machinery for retinoid processing, supporting visual pigment regeneration.
  • Demonstrated complete visual cycle activity in iPS-RPE, validating its potential for transplantation.