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iPreP is a three-dimensional nanofibrillar cellulose hydrogel platform for long-term ex vivo preservation of human islets
Yi-Ju Chen, … , Paul Gadue, Ben Z. Stanger
Yi-Ju Chen, … , Paul Gadue, Ben Z. Stanger
Published November 1, 2019
Citation Information: JCI Insight. 2019;4(21):e124644. https://doi.org/10.1172/jci.insight.124644.
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Resource and Technical Advance Endocrinology

iPreP is a three-dimensional nanofibrillar cellulose hydrogel platform for long-term ex vivo preservation of human islets

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Abstract

Islet transplantation is an effective therapy for achieving and maintaining normoglycemia in patients with type 1 diabetes mellitus. However, the supply of transplantable human islets is limited. Upon removal from the pancreas, islets rapidly disintegrate and lose function, resulting in a short interval for studies of islet biology and pretransplantation assessment. Here, we developed a biomimetic platform that can sustain human islet physiology for a prolonged period ex vivo. Our approach involved the creation of a multichannel perifusion system to monitor dynamic insulin secretion and intracellular calcium flux simultaneously, enabling the systematic evaluation of glucose-stimulated insulin secretion under multiple conditions. Using this tool, we developed a nanofibrillar cellulose hydrogel–based islet-preserving platform (iPreP) that can preserve islet viability, morphology, and function for nearly 12 weeks ex vivo, and with the ability to ameliorate glucose levels upon transplantation into diabetic hosts. Our platform has potential applications in the prolonged maintenance of human islets, providing an expanded time window for pretransplantation assessment and islet studies.

Authors

Yi-Ju Chen, Taiji Yamazoe, Karla F. Leavens, Fabian L. Cardenas-Diaz, Andrei Georgescu, Dongeun Huh, Paul Gadue, Ben Z. Stanger

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Figure 3

A 3D NFC hydrogel platform for maintaining human islet function and morphology in a long-term.

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A 3D NFC hydrogel platform for maintaining human islet function and morp...
(A) Viability assessment of human islets encapsulated in a 3D NFC hydrogel platform. Top: Schematic of the 3D NFC hydrogel platform. Bottom: Representative images of an islet after 21 days cultured in NFC hydrogel platform, followed by LIVE/DEAD viability staining and imaged for morphology (bright-field), viable cells (green fluorescence), and apoptotic cells (red fluorescence). Inset: Representative image of an islet cultured for 7 days in standard (CIT) medium, followed by LIVE/DEAD staining. Scale bar: 100 μm. (B) Human islets cultured for 14 or 21 days in NFC hydrogel with CIT media were functionally similar to their freshly isolated (day 1) counterparts. The experiments were conducted using islets from the same donors as in Figure 2. Results were obtained from 3 independent experiments. Each experiment was performed in duplicate or triplicate with islets isolated from the same donor. The number of donors used in each condition (n = 2 or n = 3) is indicated. Values are shown as mean ± SEM. Insulin values were normalized to DNA content of each sample. (C) A comparison of human islets cultured for 42 days in Transwell or with 3D NFC hydrogels. Human islets cultured in Transwell for 42 days exhibited significantly higher levels of basal insulin secretion in response to low glucose (1 mM and 3 mM) compared with fresh islets (day 1) or hydrogel-cultured islets. Experiments were conducted 4 times using samples from 4 different cadaveric donors. Each experiment was performed in duplicate or triplicate with islets isolated from the same donor. Values were shown as mean ± SEM. Insulin values were normalized to DNA content of each sample. Multiple-comparisons t test was performed, followed by Holm-Šidák correction. (D) Representative images of human islets following long-term NFC hydrogel culture. Cultured human islets were retrieved on day 14, 21, 42, or 83 by treatment with cellulase and imaged for morphology (bright-field). Scale bars: 100 μm.

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