Cell Counting and Cell Cycle Analysis of Simple Non-Cultured Endothelial Cell Injection (SNEC-I) Therapy:...

Area of Science:

• Ophthalmology
• Regenerative Medicine
• Cell Therapy

Background:

• Corneal endothelial diseases pose a significant challenge in ophthalmology.
• Current treatments often involve complex procedures or cell culture.
• Novel therapeutic strategies are needed for corneal regeneration.

Purpose of the Study:

• To develop a semi-automated cell counting method for corneal endothelial cells (CEnCs).
• To characterize the extent of cellular manipulation in SNEC-I therapy.
• To evaluate the translational workflow and efficacy of SNEC-I therapy in a preclinical model.

Main Methods:

• Comparison of manual and automated cell counting techniques.
• Analysis of CEnC cell cycle status and gene expression.
• Single-cell RNA-sequencing for transcriptomic profiling.
• Evaluation of SNEC-I therapy in a rabbit model of bullous keratopathy.

Main Results:

• Manual and automated cell counting methods demonstrated comparable accuracy and reproducibility.
• SNEC-I-harvested CEnCs showed minimal cellular manipulation, closely resembling native corneal endothelium.
• Successful resolution of bullous keratopathy in the rabbit model confirmed SNEC-I therapy's feasibility, efficacy, and safety.

Conclusions:

• SNEC-I therapy offers a robust and minimally manipulated approach for corneal endothelial regeneration.
• The findings support SNEC-I therapy as an attractive therapeutic option from a regulatory perspective.
• This study validates the translational potential of SNEC-I for treating corneal endothelial diseases.