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3D co-culturing model of primary pancreatic islets and hepatocytes in hybrid spheroid to overcome pancreatic cell shortage

Authors
Jun, YeslKang, Ah RanLee, Jae SeoJeong, Gi SeokJu, JongilLee, Dong YunLee, Sang-Hoon
Issue Date
5월-2013
Publisher
ELSEVIER SCI LTD
Keywords
Co-culture; Pancreatic islets; Hepatocytes; Concave microwell array; Type-1 diabetes mellitus (T1DM); Islet encapsulation
Citation
BIOMATERIALS, v.34, no.15, pp.3784 - 3794
Indexed
SCIE
SCOPUS
Journal Title
BIOMATERIALS
Volume
34
Number
15
Start Page
3784
End Page
3794
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/103381
DOI
10.1016/j.biomaterials.2013.02.010
ISSN
0142-9612
Abstract
Here, a spheroidal 3D co-culture model of primary (rat) pancreatic islets and hepatocytes with uniform size and shape was developed using hemispheric concave microwell arrays. We conducted morphological and functional analyses of hybrid spheroids versus mono-cultures of islets or hepatocytes (controls). For the establishment of a 3D hybrid model, a broad range of cell ratios - 1:1, 1:3, 1:5, 1:7, 3:1, 5:1 and 7:1 mixture - of hepatocytes and pancreatic islets were used. As control, each hepatocyte and pancreatic islet were mono-cultured forming 3D spheroids. The transient morphology of spheroid formation in 9 culture models was observed using optical microscopy. Cell viability under these culture environments was assessed, and the morphologies of the outer and inner porous cell-spheroid structures were investigated using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and imaging of stained spheroid sections. The pancreatic islet-specific function of hybrid spheroids was evaluated by measuring insulin secretion and in vivo test by xenotransplantation of encapsulated spheroids in microfibers with a consistent maintenance of normal blood glucose levels over 4 weeks, while liver-specific functions were measured in terms of albumin secretion, urea secretion and cytochrome P450 activity. These diverse observations and evaluations validated the positive and bidirectional effects of co-cultured 3D spheroids. The proposed 3D co-culture model demonstrated that both cells appeared to support each other's functions strongly in spheroids, even though smaller proportions of each cell type was evaluated compared to mono-culture models, suggesting that the proposed model could help overcome the problem of cell shortages in clinical applications. (C) 2013 Elsevier Ltd. All rights reserved.
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