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Updated: Jun 17, 2026
A Simple Mechanical Procedure to Create Limbal Stem Cell Deficiency in Mouse
Published on: November 17, 2016
Corneal surface reconstruction using adult mesenchymal stem cells in experimental limbal stem cell deficiency in
Helga Reinshagen1, Claudia Auw-Haedrich, Ruediger V Sorg
1Eye Center, Clinic Pallas, Olten, Switzerland. helga.reinshagen-spelsberg@klinik-pallas.ch
Purpose:
To investigate the ability of mesenchymal stem cells (MSC) to transdifferentiate to corneal epithelial cells in experimental limbal stem cell deficiency in rabbits.
Methods:
Total limbal stem cell deficiency was produced in 21 right eyes of 21 New Zealand rabbits; 6 eyes served as controls (group 1, G1). After removal of the conjunctival overgrowth, five eyes received amniotic membrane transplantation (AMT; G2). In four eyes, autologous limbal stem cell transplantation from the healthy eye was performed with AMT (G3). In another six eyes, enriched autologous MSC were injected under the amniotic membrane (AM) (G4). Within 280 days, corneoscleral discs were analysed for goblet cells, cytokeratin (CK) 3/12, connexin 43, β(1) -integrin, CK 19, α-enolase, p63 and ATP-binding cassette transporter subtype G-2 (ABCG-2) distribution patterns.
Results:
Cultivated MSC were positive for CK 3/12 and α-enolase, but negative for ABCG-2, p63 and connexin 43. On rabbit corneas, CK 3/12 was expressed in all corneal regions in all groups, but with significantly different intensities. Among all other parameters, expression levels of ABCG-2, β(1) -integrin and connexin 43 were significantly different between the transplanted groups and the control group. After a mean follow-up time of 172 (47-280) days, goblet cells were rarely present in the central cornea (G1-4).
Conclusion:
CK 3/12 is not highly specific for differentiated corneal epithelium. Further, goblet cells are not a reliable marker for conjunctivalization in rabbits. Expression of ABCG-2, β(1)-integrin and connexin 43 after mesenchymal stem cell transplantation may indicate their ability to maintain their stem cell character or to transdifferentiate to epithelial progenitor cells.
Insights
Mesenchymal stem cells (MSC) showed potential for corneal epithelial cell transdifferentiation in rabbits with limbal stem cell deficiency. Markers like ABCG-2, β(1)-integrin, and connexin 43 suggest MSCs may retain stemness or become epithelial progenitors.
Area of Science:
- Ophthalmology
- Stem Cell Biology
- Regenerative Medicine
Background:
- Limbal stem cell deficiency (LSCD) impairs corneal regeneration, leading to visual impairment.
- Mesenchymal stem cells (MSCs) are multipotent stromal cells with immunomodulatory and regenerative potential.
- Investigating MSCs' transdifferentiation capacity is crucial for developing novel LSCD therapies.
Purpose of the Study:
- To evaluate the transdifferentiation potential of mesenchymal stem cells (MSCs) into corneal epithelial cells.
- To assess the efficacy of MSC transplantation in a rabbit model of experimental limbal stem cell deficiency.
- To analyze the expression of key corneal and stem cell markers post-transplantation.
Main Methods:
- Induction of total limbal stem cell deficiency in rabbit corneas.
- Comparison of treatment groups: control, amniotic membrane transplantation (AMT), autologous limbal stem cell transplantation with AMT, and MSC injection under AM.
- Analysis of corneoscleral tissues for markers including cytokeratin 3/12, connexin 43, β(1)-integrin, and ABCG-2.
Main Results:
- Cultured MSCs expressed CK 3/12 and α-enolase but lacked ABCG-2, p63, and connexin 43.
- CK 3/12 expression varied in intensity across corneal regions and groups.
- Significantly different expression levels of ABCG-2, β(1)-integrin, and connexin 43 were observed between transplanted and control groups.
Conclusions:
- Cytokeratin 3/12 is not a definitive marker for differentiated corneal epithelium.
- Goblet cells are unreliable indicators of conjunctivalization in rabbit models.
- Post-transplantation expression of ABCG-2, β(1)-integrin, and connexin 43 suggests MSCs may maintain stemness or transdifferentiate into epithelial progenitor cells.
