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Micropyramid-patterned, oxygen-permeable bottomed dish for high density culture of pancreatic islets

Myrick, Ryan J. and Shang, Kuang-Ming and Betts, Jonathan F. and Gonzalez, Nelson and Rawson, Jeffrey and Izumi, Kenji and Koba, Naoya and Tsuchiya, Takanori and Kato, Hiroyuki and Omori, Keiko and Kandeel, Fouad and Mullen, Yoko and Tai, Yu-Chong and Botvinick, Elliot and Komatsu, Hirotake (2023) Micropyramid-patterned, oxygen-permeable bottomed dish for high density culture of pancreatic islets. Biofabrication, 15 (1). Art. No. 015018. ISSN 1758-5082. doi:10.1088/1758-5090/aca79a.

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The need for maintaining cell-spheroid viability and function within high-density cultures is unmet for various clinical and experimental applications, including cell therapies. One immediate application is for transplantation of pancreatic islets, a clinically recognized treatment option to cure type 1 diabetes; islets are isolated from a donor for subsequent culture prior to transplantation. However, high seeding conditions cause unsolicited fusion of multiple spheroids, thereby limiting oxygen diffusion to induce hypoxic cell death. Here we introduce a culture dish incorporating a micropyramid-patterned surface to prevent the unsolicited fusion and oxygen-permeable bottom for optimal oxygen environment. A 400 µm-thick, oxygen-permeable polydimethylsiloxane sheet topped with micropyramid pattern of 400 µm-base and 200 µm-height was fabricated to apply to the 24-well plate format. The micropyramid pattern separated the individual pancreatic islets to prevent the fusion of multiple islets. This platform supported the high oxygen demand of islets at high seeding density at 260 islet equivalents cm⁻², a 2–3-fold higher seeding density compared to the conventional islet culture used in a preparation for the clinical islet transplantations, demonstrating improved islet morphology, metabolism and function in a 4 d-culture. Transplantation of these islets into immunodeficient diabetic mice exhibited significantly improved engraftment to achieve euglycemia compared to islets cultured in the conventional culture wells. Collectively, this simple design modification allows for high-density cultures of three-dimensional cell spheroids to improve the viability and function for an array of investigational and clinical replacement tissues.

Item Type:Article
Related URLs:
URLURL TypeDescription
Myrick, Ryan J.0000-0001-5506-5782
Shang, Kuang-Ming0000-0001-5065-7607
Kato, Hiroyuki0000-0001-6839-7663
Kandeel, Fouad0000-0003-3746-5015
Tai, Yu-Chong0000-0001-8529-106X
Botvinick, Elliot0000-0001-9837-805X
Komatsu, Hirotake0000-0003-0876-4809
Additional Information:We thank Sung Hee Kil, Ph D for critical reading and editing of the manuscript and Juan Carreno for the equipment preparations. Eiji Kobayashi, M D, Ph D provided Luc-Tg LEW rats. Data availability statement. All data that support the findings of this study are included within the article (and any sentry files). Additional data related to this paper may be available to researchers upon request. Funding: Nora Eccles Treadwell Foundation (no Grant Number, HKo); National Institutes of Health Grant R03DK129958-01 (HKo)
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Nora Eccles Treadwell FoundationUNSPECIFIED
Issue or Number:1
Record Number:CaltechAUTHORS:20230103-817548100.28
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:118628
Deposited By: Research Services Depository
Deposited On:27 Jan 2023 18:30
Last Modified:27 Jan 2023 18:30

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