Poster #090, Cedars Sinai Medical Center
Development of a Biomimetic GelMA Corneal Model with Human Limbal Stem Cells
Mentors: Daxian Zha, PhD and Mehrnoosh Saghizadeh Ghiam, PhD
Approximately 2.2 billion people worldwide are affected by near or distance visual impairment, with corneal disease representing a major cause. A global shortage of donor corneas further worsens this problem. Replicating the cornea’s curved architecture is challenging, but GelMA (gelatin methacrylate) can be photocrosslinked into defined shapes and has tunable properties that closely mimic corneal tissue. We developed and characterized GelMA constructs encapsulating human limbal progenitor cells. Donor corneas were dissected, and limbal epithelial cells (LECs) and limbal stromal cells (LSCs) were isolated by enzymatic digestion. A PDMS mold was created using a printed cast mold, and hydrophilic surface treatment was performed using plasma activation to facilitate demolding of the constructs while preserving their structural integrity. LSCs were suspended in a filtered GelMa solution containing LAP (Lithium phenyl-2,4,6-trimethylbenzoylphosphinate), the photoinitiator, cast into the mold and photocrosslinked under UV light to form stable hydrogel constructs. The constructs remained in media where Calcium AM staining was performed every 5 days to ensure cell viability. After 4 weeks of maturation, LECs were seeded onto the constructs and cultured at an air–liquid interface to induce stratification. The constructs were immunostained to assess LEC and LSC phenotypes. Confocal imaging revealed localization of the cells into their native corneal architecture, with LECs oriented on the top, outer layer of the construct, and the LSCs oriented in the inner layer. Our findings suggest that the GelMA constructs not only allow for cell proliferation, but that they can also mimic the micro-environments and phenotypes of the cornea, showing that the cells in the GelMA construct offers stromal and epithelial cell interactions. Overall, the GelMA constructs provide a clear pathway to further research on corneal disease mechanisms and tissue regeneration.