Transcriptomic Landscape and Functional Characterization of Human Induced Pluripotent Stem Cell-Derived Limbal...

Area of Science:

• Ophthalmology
• Regenerative Medicine
• Stem Cell Biology

Background:

• Limbal stem cell deficiency (LSCD) impairs corneal health and transparency by affecting limbal epithelial progenitor cells (LEPC).
• Human induced pluripotent stem cell-derived LEPC (hiPSC-LEPC) offer a potential therapeutic avenue for LSCD.
• Detailed comparison of hiPSC-LEPC with tissue-derived LEPC (T-LEPC) is needed to assess their therapeutic potential.

Purpose of the Study:

• To compare the characteristics and functions of hiPSC-LEPC with T-LEPC.
• To evaluate the potential of hiPSC-LEPC for corneal regeneration in vitro.
• To identify challenges for clinical application of hiPSC-LEPC in LSCD treatment.

Main Methods:

• Gene expression profiling of hiPSC-LEPC and T-LEPC.
• In vitro assays for colony-forming ability and wound healing.
• Immunogenicity assessment via peripheral blood mononuclear cell proliferation assays.
• Evaluation of hiPSC-LEPC and their secretome on vascular endothelial cell proliferation.
• Repopulation of decellularized human corneolimbal (DHC/L) scaffolds with hiPSC-LEPC.

Main Results:

• hiPSC-LEPC and T-LEPC exhibited similar gene expression, colony formation, wound healing, and melanosome uptake.
• hiPSC-LEPC demonstrated lower immunogenicity and reduced peripheral blood mononuclear cell proliferation compared to T-LEPC.
• The secretome of hiPSC-LEPC was more effective at inhibiting vascular endothelial cell proliferation than that of T-LEPC.
• hiPSC-LEPC successfully formed multilayered epithelium on DHC/L scaffolds, with some abnormal basal extracellular matrix deposition observed.

Conclusions:

• hiPSC-LEPC possess functional properties comparable to native LEPC, suggesting their therapeutic viability for LSCD.
• hiPSC-LEPC combined with DHC/L scaffolds show potential for future corneal transplantation.
• Further research is required to address challenges like abnormal extracellular matrix deposition before clinical implementation.