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Reduction of Activin A gives rise to comparable expression of key definitive endoderm and mature beta cell markers

Aldyn Wildey

Stephen Harrington

Lisa Stehno-Bittel

Francis Karanu

Aldyn Wildey

Likarda LLC, Kansas City, MO 64137, USA

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Stephen Harrington

Likarda LLC, Kansas City, MO 64137, USA

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Lisa Stehno-Bittel

Likarda LLC, Kansas City, MO 64137, USA

University of Kansas Medical Center, Kansas City, KS, USA

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Francis Karanu

*Author for correspondence:

E-mail Address: fkaranu@likarda.com

https://orcid.org/0000-0001-6235-9586

Likarda LLC, Kansas City, MO 64137, USA

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Published Online:19 Jan 2024https://doi.org/10.2217/rme-2023-0187

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Abstract

Aim: Cell therapies for diabetes rely on differentiation of stem cells into insulin-producing cells, which is complex and expensive. Our goal was to evaluate production costs and test ways to reduce it. Methods: Cost of Goods (COGs) analysis for differentiation was completed and the effects of replacement or reduction of the most expensive item was tested using qRT-PCR, immunohistochemistry, flow cytometry along with glucose-stimulated insulin release. Results: Activin A (AA) was responsible for significant cost. Replacement with small molecules failed to form definitive endoderm (DE). Reducing AA by 50% did not negatively affect expression of beta cell markers. Conclusion: Reduction of AA concentration is feasible without adversely affecting DE and islet-like cell differentiation, leading to significant cost savings in manufacturing.

Keywords:

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Vol. 19, No. 1

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History

Received 20 September 2023

Accepted 5 January 2024

Published online 19 January 2024

Published in print January 2024

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Keywords

Author contributions

A Wildey: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing and review, final approval of manuscript. S Harrington: data analysis and interpretation, manuscript writing and review, final approval of manuscript. L Stehno-Bittel: data analysis and interpretation, financial support, administrative support, manuscript writing and review, final approval of manuscript. F Karanu: conception and design, data analysis and interpretation, manuscript writing and review, final approval of manuscript.

Acknowledgments

We wish to thank S Billingsly for assistance with qRT-PCR analyses and Karthik Ramachandran for discussions concerning cost of goods. We also thank D Muathe for help with flow cytometry and general lab support.

Financial disclosure

This study was fully supported and funded by Likarda, LLC. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

Authors are employees of Likarda, LLC. Lisa Stehno-Bittel is a founder and has equity in Likarda. The authors have no other competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Open access

This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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