Human-Induced Pluripotent Stem Cells Generate Light Responsive Retinal Organoids with Variable and Nutrient-Dependent...

Area of Science:

• Stem Cell Biology
• Retinal Development
• Ophthalmology

Background:

• Developing in vitro models of the human retina is essential for pharmacological and toxicological research.
• Generating laminated, functional, and light-responsive retinal organoids from patient-specific induced pluripotent stem cells (iPSC) is a key challenge.

Purpose of the Study:

• To investigate the efficiency of different human iPSC lines in generating retinal organoids.
• To characterize the physiological function and light responsiveness of these organoids.
• To optimize the differentiation protocol for scalability and drug screening.

Main Methods:

• Investigated five human iPSC lines for retinal organoid differentiation.
• Utilized a factorial experimental design to optimize seeding density and nutrient availability.
• Adapted the protocol to a multiwell plate format for enhanced generation.
• Tested the response of organoids to Moxifloxacin.

Main Results:

• Significant variability was observed in retinal organoid generation efficiency among iPSC lines.
• By 5 months, all organoids exhibited light responses comparable to neonatal mouse retina.
• Organoids developed an outer nuclear-like layer with photoreceptors and light-sensing structures.
• Optimized protocol improved retinal-pigmented epithelium and ganglion cell development.
• Moxifloxacin primarily affected photoreceptor cells, similar to in vivo findings.

Conclusions:

• Carefully selected iPSC lines can efficiently generate light-responsive retinal organoids.
• These organoids are suitable for large-scale pharmacological and drug screening applications.
• In vitro models accurately reflect in vivo drug responses in retinal cells.