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Related Experiment Video

Updated: Dec 6, 2025

Characterization of a Novel Human Organotypic Retinal Culture Technique

Characterization of a Novel Human Organotypic Retinal Culture Technique

Published on: June 9, 2021

Semi-Automated Approach for Retinal Tissue Differentiation.

Evgenii Kegeles^1,2, Tatiana Perepelkina¹, Petr Baranov¹

¹The Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA.

Translational Vision Science & Technology

|October 7, 2020

Summary

Scaling up retinal organoid differentiation using a semiautomated approach significantly reduces hands-on time and enables efficient analysis. This method enhances throughput for stem cell-derived retinal tissue production and research.

Keywords:

automation forskolin mouse embryonic stem cells organoids pluripotent cells retina

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Last Updated: Dec 6, 2025

Characterization of a Novel Human Organotypic Retinal Culture Technique

Characterization of a Novel Human Organotypic Retinal Culture Technique

Published on: June 9, 2021

En face Cryosectioning of Mouse Retina for High-dimensional Spatial Molecular Analysis

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Published on: July 8, 2025

Efficient and Consistent Generation of Retinal Pigment Epithelium/Choroid Flatmounts from Human Eyes for Histological Analysis

Efficient and Consistent Generation of Retinal Pigment Epithelium/Choroid Flatmounts from Human Eyes for Histological Analysis

Published on: October 28, 2022

Area of Science:

Stem cell biology
Retinal development
Biotechnology

Background:

Three-dimensional differentiation of pluripotent stem cells is crucial for studying retinal development.
Current protocols face limitations in throughput for cell production and drug discovery.

Purpose of the Study:

To scale up a three-dimensional retinal differentiation protocol using a semiautomated approach.
To improve the efficiency and throughput of retinal organoid production.

Main Methods:

Utilized mouse embryonic stem cells (Rx-GFP) and human embryonic stem cells (Brn3b-tdTomato).
Implemented automated media exchange with robotic systems (Thermo WellWash Versa, Thermo RapidStack).
Employed automated image analysis with custom Python OpenCV scripts for differentiation assessment.

Main Results:

Semiautomated approach reduced operator time from two hours to 34 minutes for 960 organoids over 25 days.
Differentiation pattern and quantity remained unchanged.
Automated image analysis revealed Forskolin treatment significantly increased retinal field induction efficiency.

Conclusions:

Semiautomated methods increase throughput for retinal tissue differentiation protocols.
Automated image analysis is effective for evaluating differentiation efficiency and identifying factors influencing it.
Robotic approaches minimize human error and support enclosed cell production for GMP manufacturing.