Advances in pancreatic islet monolayer culture on glass surfaces enable super-resolution microscopy and insights into beta cell ciliogenesis and proliferation
| Autor: | Jaime Santo-Domingo, Andreas Wiederkehr, Gabriele Galliverti, Jeffrey A. Hubbell, Olivier Burri, Steinunn Baekkeskov, Lorenzo Piemonti, Chiara Cianciaruso, Edward A. Phelps, Ekaterine Berishvili, Miriella Pasquier |
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| Přispěvatelé: | Pathology/molecular and cellular medicine, Phelps, E. A., Cianciaruso, C., Santo Domingo, J., Pasquier, M., Galliverti, G., Piemonti, Lorenzo, Berishvili, E., Burri, O., Wiederkehr, A., Hubbell, J. A., Baekkeskov, S. |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Male Glass/chemistry Cellular differentiation Organogenesis Cell Cell Culture Techniques Cell Culture Techniques/methods Hippocampus Microtubules Rats Sprague-Dawley Cilia/metabolism Insulin-Secreting Cells Glucose/metabolism Insulin Cells Cultured Neurons Microscopy Multidisciplinary geography.geographical_feature_category Cultured ddc:617 Diabetes Cell Differentiation Middle Aged Islet Cell biology medicine.anatomical_structure Phenotype Microtubules/metabolism Actins/metabolism Hippocampus/cytology Insulin-Secreting Cells/cytology/metabolism Female Beta cell Microscopy/methods Biotechnology Adult endocrine system Calcium/metabolism Insulin/metabolism Adolescent Cells 1.1 Normal biological development and functioning Bioengineering Biology 03 medical and health sciences Young Adult Underpinning research Ciliogenesis Journal Article medicine Cell Adhesion Animals Humans Cilia Cell adhesion Cell Proliferation geography Cell growth In vitro Actins Coculture Techniques Rats 030104 developmental biology Glucose Calcium Sprague-Dawley Glass Digestive Diseases Neurons/cytology/metabolism |
| Zdroj: | Scientific reports, vol 7, iss 1 Scientific Reports, Vol. 7 (2017) P. 45961 |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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