Abstract
Purpose :
Corneal wounding and limbal stem cell deficiency (LSCD) can lead to significant visual impairment and ocular health challenges. Tissue engineering approaches using stem cells seeded on biomaterials show promise for corneal repair and offer a potential solution to restore vision and improve patient outcomes. Autologous adult stem cells isolated from various tissues such as bone marrow, fat, and corneal-limbal stroma have multipotent potential to promote corneal repair. However, obtaining these stem cells requires invasive biopsies. This study investigates the regenerative potential of human urine-derived stem cells (USCs), a non-invasive and readily available stem cell population, seeded on decellularized small intestinal submucosa (SIS) to generate autologous stratified epithelium for corneal repair.
Methods :
USCs were isolated from healthy donors (n=9, M:F 6:3). A novel differentiation protocol was established to direct USCs towards a corneal epithelial lineage by evaluating epithelial cell markers, tight junction formation via transwell assays, and barrier function after epithelial differentiation. thin SIS scaffolds seeded with USCs as an alternative corneal construct were evaluated by immunohistochemistry.
Results :
After differentiation, USCs exhibited a cobblestone morphology, a multilayered well-stratified epithelium, and microvilli-studded tight junctions. Differentiated USCs expressed epithelial cell markers (cytokeratin 7, 13, 19, and 20). The 3-5 layered epithelium of USCs showed appropriate structural and functional characteristics resembling native corneal epithelium as evidenced by tight junction proteins (ZO-1, ZO-2, claudins, and occludin) and barrier functional characteristics. The stratified epithelium of SIS seeded with differentiated USCs is positive for pan cytokeratin AE1/AE3.
Conclusions :
The results suggest that human USCs have significant potential as a convenient, low-cost, and personalized source for the generation of autologous corneal epithelial constructs for the treatment of corneal defects and LSCD. This approach opens new avenues for personalized regenerative medicine in ophthalmology and may pave the way for innovative treatments of various corneal diseases and injuries, corneal disease modeling, and drug testing.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.