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Title

Advances in pancreatic islet monolayer culture on glass surfaces enable super-resolution microscopy and insights into beta cell ciliogenesis and proliferation

Authors
Phelps, Edward A
Cianciaruso, Chiara
Santo-Domingo, Jaime
Pasquier, Miriella
Galliverti, Gabriele
Piemonti, Lorenzo
Burri, Olivier
show hidden authors show all authors [1 - 11]
Published in Scientific reports. 2017, vol. 7, 45961
Abstract A robust and reproducible method for culturing monolayers of adherent and well-spread primary islet cells on glass coverslips is required for detailed imaging studies by super-resolution and live-cell microscopy. Guided by an observation that dispersed islet cells spread and adhere well on glass surfaces in neuronal co-culture and form a monolayer of connected cells, we demonstrate that in the absence of neurons, well-defined surface coatings combined with components of neuronal culture media collectively support robust attachment and growth of primary human or rat islet cells as monolayers on glass surfaces. The islet cell monolayer cultures on glass stably maintain distinct mono-hormonal insulin+, glucagon+, somatostatin+ and PP+ cells and glucose-responsive synchronized calcium signaling as well as expression of the transcription factors Pdx-1 and NKX-6.1 in beta cells. This technical advance enabled detailed observation of sub-cellular processes in primary human and rat beta cells by super-resolution microscopy. The protocol is envisaged to have broad applicability to sophisticated analyses of pancreatic islet cells that reveal new biological insights, as demonstrated by the identification of an in vitro protocol that markedly increases proliferation of primary beta cells and is associated with a reduction in ciliated, ostensibly proliferation-suppressed beta cells.
Keywords Actins/metabolismAdolescentAdultAnimalsCalcium/metabolismCell AdhesionCell Culture Techniques/methodsCell DifferentiationCell ProliferationCells, CulturedCilia/metabolismCoculture TechniquesFemaleGlass/chemistryGlucose/metabolismHippocampus/cytologyHumansInsulin/metabolismInsulin-Secreting Cells/cytology/metabolismMaleMicroscopy/methodsMicrotubules/metabolismMiddle AgedNeurons/cytology/metabolismOrganogenesisPhenotypeRats, Sprague-DawleyYoung Adult
Identifiers
PMID: 28401888
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Structures
Research groups Création de tissus par bio-ingénieurie et régénération d'organes (1021)
La transplantation d'îlots de Langerhans (623)
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PHELPS, Edward A et al. Advances in pancreatic islet monolayer culture on glass surfaces enable super-resolution microscopy and insights into beta cell ciliogenesis and proliferation. In: Scientific Reports, 2017, vol. 7, p. 45961. doi: 10.1038/srep45961 https://archive-ouverte.unige.ch/unige:138362

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Deposited on : 2020-07-10

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