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Generation and Proteome Profiling of PBMC-Originated, iPSC-Derived Corneal Endothelial Cells

Muhammad Ali1, Shahid Y Khan1, Shivakumar Vasanth1

  • 1The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States.

Investigative Ophthalmology & Visual Science
|May 31, 2018
PubMed

Insights

Researchers developed corneal endothelial cells (CECs) from peripheral blood mononuclear cells (PBMCs) using induced pluripotent stem cells (iPSCs). These iPSC-derived CECs mimic human corneal endothelium at the molecular level, offering a promising personalized approach.

Area of Science:

  • Ophthalmology
  • Stem Cell Biology
  • Regenerative Medicine

Background:

  • Corneal clarity depends on corneal endothelial cells (CECs).
  • Generating functional CECs for therapeutic use remains a challenge.
  • Induced pluripotent stem cells (iPSCs) offer a potential source for cell replacement therapies.

Purpose of the Study:

  • To develop corneal endothelial cells (CECs) derived from peripheral blood mononuclear cells (PBMCs) via induced pluripotent stem cells (iPSCs).
  • To characterize the morphology, marker expression, and proteomic profile of these iPSC-derived CECs.
  • To compare the molecular architecture of iPSC-derived CECs with human corneal endothelium (hCE).

Main Methods:

  • Peripheral blood mononuclear cells (PBMCs) were isolated and reprogrammed into iPSCs.
  • iPSCs were differentiated into CECs through neural crest cells (NCCs).
  • Morphology was assessed by phase contrast microscopy, marker expression by qRT-PCR, and proteomic profiling by mass spectrometry.

Main Results:

  • PBMC-originated, iPSC-derived CECs exhibited characteristic hexagonal shapes and tight adherence.
  • CE-associated markers were significantly upregulated, while pluripotency markers were minimally expressed.
  • Proteomic analysis revealed a high overlap (90.82%) between iPSC-derived CECs and hCE proteomes.

Conclusions:

  • Successfully developed a personalized method for generating iPSC-derived CECs from PBMCs.
  • These iPSC-derived CECs closely mimic the molecular architecture of native hCE.
  • This represents the first report of PBMC-originated, iPSC-derived CECs, paving the way for potential therapeutic applications.
Abstract

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