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Innovative Microcapsules for Pancreatic β-Cells Harvested from Mature Double-Transgenic Mice: Cell Imaging, Viability, Induced Glucose-Stimulated Insulin Measurements and Proinflammatory Cytokines Analysis

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Abstract

Purpose

Recently we demonstrated that microencapsulation of a murine pancreatic β-cell line using an alginate-ursodeoxycholic acid (UDCA) matrix produced microcapsules with good stability and cell viability. In this study, we investigated if translation of this formulation to microencapsulation of primary β-cells harvested from mature double-transgenic healthy mice would also generate stable microcapsules with good cell viability.

Methods

Islets of Langerhans were isolated from Ngn3-GFP/RIP-DsRED mice by intraductal collagenase P digestion and density gradient centrifugation, dissociated into single cells and the β-cell population purified by Fluorescence Activated Cell Sorting. β-cells were microencapsulated using either alginate-poly-l-ornithine (F1; control) or alginate-poly-l-ornithine-UDCA (F2; test) formulations. Microcapsules were microscopically examined and microencapsulated cells were analyzed for viability, insulin and cytokine release, 2 days post-microencapsulation.

Results

Microcapsules showed good uniformity and morphological characteristics and even cell distribution within microcapsules with or without UDCA. Two days post microencapsulation cell viability, mitochondrial ATP and insulin production were shown to be optimized in the presence of UDCA whilst production of the proinflammatory cytokine IL-1β was reduced. Contradictory to our previous studies, UDCA did not reduce production of any other pro-inflammatory biomarkers.

Conclusions

These results suggest that UDCA incorporation improves microcapsules’ physical and morphological characteristics and improves the viability and function of encapsulated mature primary pancreatic β-cells.

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Abbreviations

ATP:

Adenosine triphosphate

CMRL:

Connaught Medical Research Laboratories

DAPI:

Diamidino-2-phenylindole

FCS:

Fetal calf serum

IFN-γ:

Interferon-γ

IL-1β:

Interleukin-1β

IL-6:

Interleukin-6

IL-12:

Interleukin-12

IL-17:

Interleukin-17

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H–tetrazolium

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

Ngn3-GFP:

Neurogenin3-Green fluorescent protein

PBS:

Phosphate buffered saline

PLO:

Poly-l-ornithine

RIP-DsRED:

Rat insulin promoter-Discosoma sp. red fluorescent protein

SA:

Sodium alginate

TNF-α:

Tumor necrosis factor-α

UDCA:

Ursodeoxycholic acid

XTT:

2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H–Tetrazolium-5-Carboxanilide

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Authors and Affiliations

  1. Biotechnology and Drug Development Research Laboratory, School of Pharmacy, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Perth, WA, Australia

    Armin Mooranian & Hani Al-Salami

  2. School of Public Health, Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia

    Ryu Tackechi

  3. Harry Perkins Institute of Medical Research, Centre for Diabetes Research, Perth, WA, Australia

    Emma Jamieson & Grant Morahan

Authors
  1. Armin Mooranian
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  2. Ryu Tackechi
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  3. Emma Jamieson
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  4. Grant Morahan
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  5. Hani Al-Salami
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Correspondence to Hani Al-Salami.

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Mooranian, A., Tackechi, R., Jamieson, E. et al. Innovative Microcapsules for Pancreatic β-Cells Harvested from Mature Double-Transgenic Mice: Cell Imaging, Viability, Induced Glucose-Stimulated Insulin Measurements and Proinflammatory Cytokines Analysis. Pharm Res 34, 1217–1223 (2017). https://doi.org/10.1007/s11095-017-2138-y

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  • DOI: https://doi.org/10.1007/s11095-017-2138-y

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