Automating iPSC generation to enable autologous photoreceptor cell replacement therapy

Area of Science:

• Regenerative Medicine
• Stem Cell Biology
• Ophthalmology

Background:

• Inherited retinal degeneration causes incurable vision loss.
• Autologous induced pluripotent stem cell (iPSC)-mediated photoreceptor cell replacement is a potential therapy.
• Lack of high-throughput technologies hinders clinical translation of patient-specific iPSC therapies.

Purpose of the Study:

• To describe a robotic cell culture platform for parallel production of clinical-grade patient-specific iPSCs.
• To demonstrate the platform's capability for generating iPSCs and retinal organoids.
• To establish standard operating procedures for cGMP-compliant iPSC generation.

Main Methods:

• Utilized the Cell X precision robotic cell culture platform within an ISO Class 5 cGMP-compliant isolator.
• Performed all procedures, from fibroblast culture to iPSC generation, clonal expansion, and retinal differentiation, robotically.
• Employed scorecard analysis, karyotyping, immunostaining, confocal microscopy, and single-cell RNA sequencing for characterization.

Main Results:

• Generated patient-specific iPSCs confirmed as pluripotent and genetically stable.
• Produced retinal organoids indistinguishable from manually generated ones.
• Single-cell RNA sequencing showed comparable cell quality to manually generated iPSCs.

Conclusions:

• Successfully developed a robotic platform and SOPs for producing clinical-grade photoreceptor precursor cells.
• The system adheres to current good manufacturing practices (cGMP).
• This robotic platform enables clinical-grade iPSC production for autologous retinal cell replacement therapies.