Toward Corneal Limbus In Vitro Model: Regulation of hPSC-LSC Phenotype by Matrix Stiffness and Topography During Cell...

Area of Science:

• Ophthalmology
• Stem Cell Biology
• Biomaterials Science

Background:

• A functional limbal epithelial stem cell (LSC) niche is essential for corneal epithelium renewal and vision maintenance.
• Limited knowledge exists regarding the niche's structural and mechanical influence on LSC regulation, hindering LSC-based therapy development.

Purpose of the Study:

• To investigate the impact of mechanical and architectural niche properties on the phenotype and growth of human pluripotent stem cell-derived LSCs (hPSC-LSCs) in vitro.
• To explore the role of substrate stiffness and topography in regulating hPSC-LSC behavior.

Main Methods:

• Culture and characterization of hPSC-LSCs on polyacrylamide gels with varying stiffnesses.
• Utilized limbal-mimicking topography in polydimethylsiloxane for early-stage hPSC-LSC culture.
• Analyzed key corneal cell protein expression in native human donor corneas via whole mount staining for comparison.

Main Results:

• hPSC-LSC mechanical response and substrate preference are significantly dependent on their developmental stage.
• Evidence suggests cells may retain a 'mechanical memory' from prior culture matrices.
• The study highlights the importance of mechanical design in functional in vitro limbus models.

Conclusions:

• Mechanical and architectural cues of the LSC niche play a critical role in regulating hPSC-LSC behavior.
• In vitro models must carefully consider substrate properties to accurately mimic the in vivo LSC niche.
• These findings advance the development of LSC-based regenerative therapies for corneal diseases.