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Translational Vision Science and Technology, volume 9, issue 10, pages 24

Semi-Automated Approach for Retinal Tissue Differentiation

Kegeles Evgenii ^{1, 2}

Perepelkina Tatiana ¹

Baranov Petr ¹

Hide authors affiliations Show authors affiliations: 2 affiliations

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

Moscow Institute of Physics and Technology, Dolgoprudniy, Russia |

Publication type: Journal Article

Publication date: 2020-09-23

Association for Research in Vision and Ophthalmology (ARVO)

Journal: Translational Vision Science and Technology

Quartile SCImago

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Impact factor: 3

ISSN: 21642591

DOI: 10.1167/tvst.9.10.24

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PubMed ID: 33024617

Biomedical Engineering

Ophthalmology

Abstract

Purpose Three-dimensional strategy for the differentiation of pluripotent stem cells to the retina has been widely used to study retinal development, although the cell production and drug discovery applications are limited by the throughput. Here we attempted to scale up the protocol using a semiautomated approach. Methods For the experiments we used the Rx-GFP mouse embryonic stem cell (mES) reporter cell line, specific for early retinal development and human embryonic stem cell line Brn3b-tdTomato, specific for retinal ganglion cells. To increase the throughput, we implemented automated media exchange using Thermo WellWash Versa with Thermo RapidStack robot. To analyze the rate of retinal differentiation in mouse stem-cell derived organoids we imaged the plates at day 10 of differentiation using Life Technologies EVOS Fl Auto. The automated image analysis of fluorescent images was performed with custom Python OpenCV script. Results The implementation of a semiautomated approach significantly reduced the operator time needed: 34 minutes versus two hours for 960 organoids over the course of 25 days without any change in differentiation pattern and quantity of retinal differentiation. Automated image analysis showed that Forskolin treatment starting from day 1 leads to a significant increase in retinal field induction efficiency. Conclusions Semiautomated approach can be applied to retinal tissue differentiation to increase the throughput of the protocol. We demonstrated that automated image analysis can be used to evaluate differentiation efficiency, as well as for troubleshooting and to study factors affecting retinal differentiation. Translational Relevance Using robotic approach reduces the risk of human error and allows to perform all cycle of cell production in enclosed conditions, which is critical for GMP cell manufacture.

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Citations by journals

	1
Molecular Therapy - Methods and Clinical Development	Molecular Therapy - Methods and Clinical Development, 1, 16.67% Molecular Therapy - Methods and Clinical Development 1 publication, 16.67%
International Journal of Molecular Sciences	International Journal of Molecular Sciences, 1, 16.67% International Journal of Molecular Sciences 1 publication, 16.67%
Frontiers in Cellular Neuroscience	Frontiers in Cellular Neuroscience, 1, 16.67% Frontiers in Cellular Neuroscience 1 publication, 16.67%
SLAS Discovery	SLAS Discovery, 1, 16.67% SLAS Discovery 1 publication, 16.67%
Proceedings of the National Academy of Sciences of the United States of America	Proceedings of the National Academy of Sciences of the United States of America, 1, 16.67% Proceedings of the National Academy of Sciences of the United States of America 1 publication, 16.67%
	1

Citations by publishers

	1
Elsevier	Elsevier, 1, 16.67% Elsevier 1 publication, 16.67%
Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 1, 16.67% Multidisciplinary Digital Publishing Institute (MDPI) 1 publication, 16.67%
Frontiers Media S.A.	Frontiers Media S.A., 1, 16.67% Frontiers Media S.A. 1 publication, 16.67%
SAGE	SAGE, 1, 16.67% SAGE 1 publication, 16.67%
Proceedings of the National Academy of Sciences (PNAS)	Proceedings of the National Academy of Sciences (PNAS), 1, 16.67% Proceedings of the National Academy of Sciences (PNAS) 1 publication, 16.67%
	1

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Perepelkina T. et al. Semi-Automated Approach for Retinal Tissue Differentiation // Translational Vision Science and Technology. 2020. Vol. 9. No. 10. p. 24.

GOST all authors (up to 50) Copy

Perepelkina T., Kegeles E., Baranov P. Semi-Automated Approach for Retinal Tissue Differentiation // Translational Vision Science and Technology. 2020. Vol. 9. No. 10. p. 24.

RIS |

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RIS Copy

TY - JOUR

DO - 10.1167/tvst.9.10.24

UR - https://doi.org/10.1167%2Ftvst.9.10.24

TI - Semi-Automated Approach for Retinal Tissue Differentiation

T2 - Translational Vision Science and Technology

AU - Perepelkina, Tatiana

AU - Kegeles, Evgenii

AU - Baranov, Petr

PY - 2020

DA - 2020/09/23 00:00:00

PB - Association for Research in Vision and Ophthalmology (ARVO)

SP - 24

IS - 10

VL - 9

PMID - 33024617

SN - 2164-2591

ER -

BibTex |

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BibTex Copy

@article{2020_Perepelkina,

author = {Tatiana Perepelkina and Evgenii Kegeles and Petr Baranov},

title = {Semi-Automated Approach for Retinal Tissue Differentiation},

journal = {Translational Vision Science and Technology},

year = {2020},

volume = {9},

publisher = {Association for Research in Vision and Ophthalmology (ARVO)},

month = {sep},

url = {https://doi.org/10.1167%2Ftvst.9.10.24},

number = {10},

pages = {24},

doi = {10.1167/tvst.9.10.24}

}

MLA

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MLA Copy

Perepelkina, Tatiana, et al. “Semi-Automated Approach for Retinal Tissue Differentiation.” Translational Vision Science and Technology, vol. 9, no. 10, Sep. 2020, p. 24. https://doi.org/10.1167%2Ftvst.9.10.24.

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Publisher

Association for Research in Vision and Ophthalmology (ARVO)

Journal

Translational Vision Science and Technology

Quartile SCImago

Quartile WOS

Impact factor

ISSN

21642591 (Electronic)

Labs

MIPT Laboratory of Genomic Engineering

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