Journal article
Feeder-free differentiation of cells exhibiting characteristics of corneal endothelium from human induced pluripotent stem cells
Biology Open, Vol.7(5), pp.bio032102-bio032102
2018
DOI: 10.1242/bio.032102
PMCID: PMC5992532
PMID: 29685994
Abstract
The purpose of this study was to devise a strategy for the derivation of corneal endothelial cells (CEnCs) from adult fibroblast-derived induced pluripotent stem cells (iPSCs). IPSCs were generated from an adult human with normal ocular history via expression of OCT4 , SOX2 , KLF4 and c-MYC . Neural crest cells (NCCs) were differentiated from iPSCs via addition of CHIR99021 and SB4315542. NCCs were driven toward a CEnC fate via addition of B27, PDGF-BB and DKK-2 to CEnC media. Differentiation of NCCs and CEnCs was evaluated via rt-PCR, morphological and immunocytochemical analysis. At 17 days post-NCC induction, there were notable changes in cell morphology and upregulation of the neural crest lineage transcripts PAX3 , SOX9 , TFAP2A , SOX10 and p75NTR and the proteins p75/NGFR and SOX10. Exposure of NCCs to B27, PDGF-BB and DKK-2 induced a shift in morphology from a spindle-shaped neural phenotype to a tightly-packed hexagonal appearance and increased expression of the transcripts ATP1A1 , COL8A1 , COL8A2 , AQP1 and CDH2 and the proteins ZO-1, N-Cad, AQP-1 and Na + /K + ATPase. Replacement of NCC media with CEnC media on day 3, 5 or 8 reduced the differentiation time needed to yield CEnCs. IPSC-derived CEnCs could be used for evaluation of cornea endothelial disease pathophysiology and for testing of novel therapeutics. Summary: This study demonstrates differentiation of adult dermal fibroblast-derived iPSCs into neural crest and corneal endothelial-like cells, which could be a valuable tool for investigating the mechanisms of cornea endothelial disease.
Details
- Title: Subtitle
- Feeder-free differentiation of cells exhibiting characteristics of corneal endothelium from human induced pluripotent stem cells
- Creators
- Michael D Wagoner - ,Laura R Bohrer - ,Benjamin T Aldrich - ,Mark A Greiner - ,Robert F Mullins - ,Kristan S Worthington - ,Budd A Tucker - ,Luke A Wiley - ,
- Resource Type
- Journal article
- Publication Details
- Biology Open, Vol.7(5), pp.bio032102-bio032102
- DOI
- 10.1242/bio.032102
- PMID
- 29685994
- PMCID
- PMC5992532
- NLM abbreviation
- Biol Open
- ISSN
- 2046-6390
- eISSN
- 2046-6390
- Publisher
- The Company of Biologists Ltd
- Grant note
- ; P30 EY-025580 / ;
- Language
- English
- Date published
- 2018
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; Chemical and Biochemical Engineering; Ophthalmology and Visual Sciences
- Record Identifier
- 9983980072702771
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