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Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study

Three-dimensional (3D) cell culture by engineering spheroids has gained increasing attention in recent years because of the potential advantages of such systems over conventional two-dimensional (2D) tissue culture. Benefits include the ability of 3D to provide a more physiologically relevant enviro...

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Autores principales: Wassmer, Charles-Henri, Bellofatto, Kevin, Perez, Lisa, Lavallard, Vanessa, Cottet-Dumoulin, David, Ljubicic, Sanda, Parnaud, Géraldine, Bosco, Domenico, Berishvili, Ekaterine, Lebreton, Fanny
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7563811/
https://www.ncbi.nlm.nih.gov/pubmed/32749168
http://dx.doi.org/10.1177/0963689720937292
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author Wassmer, Charles-Henri
Bellofatto, Kevin
Perez, Lisa
Lavallard, Vanessa
Cottet-Dumoulin, David
Ljubicic, Sanda
Parnaud, Géraldine
Bosco, Domenico
Berishvili, Ekaterine
Lebreton, Fanny
author_facet Wassmer, Charles-Henri
Bellofatto, Kevin
Perez, Lisa
Lavallard, Vanessa
Cottet-Dumoulin, David
Ljubicic, Sanda
Parnaud, Géraldine
Bosco, Domenico
Berishvili, Ekaterine
Lebreton, Fanny
author_sort Wassmer, Charles-Henri
collection PubMed
description Three-dimensional (3D) cell culture by engineering spheroids has gained increasing attention in recent years because of the potential advantages of such systems over conventional two-dimensional (2D) tissue culture. Benefits include the ability of 3D to provide a more physiologically relevant environment, for the generation of uniform, size-controlled spheroids with organ-like microarchitecture and morphology. In recent years, different techniques have been described for the generation of cellular spheroids. Here, we have compared the efficiency of four different methods of islet cell aggregation. Rat pancreatic islets were dissociated into single cells before reaggregation. Spheroids were generated either by (i) self-aggregation in nonadherent petri dishes, (ii) in 3D hanging drop culture, (iii) in agarose microwell plates or (iv) using the Sphericalplate 5D™. Generated spheroids consisted of 250 cells, except for the self-aggregation method, where the number of cells per spheroid cannot be controlled. Cell function and morphology were assessed by glucose stimulated insulin secretion (GSIS) test and histology, respectively. The quantity of material, labor intensity, and time necessary for spheroid production were compared between the different techniques. Results were also compared with native islets. Native islets and self-aggregated spheroids showed an important heterogeneity in terms of size and shape and were larger than spheroids generated with the other methods. Spheroids generated in hanging drops, in the Sphericalplate 5D™, and in agarose microwell plates were homogeneous, with well-defined round shape and a mean diameter of 90 µm. GSIS results showed improved insulin secretion in response to glucose in comparison with native islets and self-aggregated spheroids. Spheroids can be generated using different techniques and each of them present advantages and inconveniences. For islet cell aggregation, we recommend, based on our results, to use the hanging drop technique, the agarose microwell plates, or the Sphericalplate 5D™ depending on the experiments, the latter being the only option available for large-scale spheroids production.
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spelling pubmed-75638112020-10-26 Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study Wassmer, Charles-Henri Bellofatto, Kevin Perez, Lisa Lavallard, Vanessa Cottet-Dumoulin, David Ljubicic, Sanda Parnaud, Géraldine Bosco, Domenico Berishvili, Ekaterine Lebreton, Fanny Cell Transplant Original Article Three-dimensional (3D) cell culture by engineering spheroids has gained increasing attention in recent years because of the potential advantages of such systems over conventional two-dimensional (2D) tissue culture. Benefits include the ability of 3D to provide a more physiologically relevant environment, for the generation of uniform, size-controlled spheroids with organ-like microarchitecture and morphology. In recent years, different techniques have been described for the generation of cellular spheroids. Here, we have compared the efficiency of four different methods of islet cell aggregation. Rat pancreatic islets were dissociated into single cells before reaggregation. Spheroids were generated either by (i) self-aggregation in nonadherent petri dishes, (ii) in 3D hanging drop culture, (iii) in agarose microwell plates or (iv) using the Sphericalplate 5D™. Generated spheroids consisted of 250 cells, except for the self-aggregation method, where the number of cells per spheroid cannot be controlled. Cell function and morphology were assessed by glucose stimulated insulin secretion (GSIS) test and histology, respectively. The quantity of material, labor intensity, and time necessary for spheroid production were compared between the different techniques. Results were also compared with native islets. Native islets and self-aggregated spheroids showed an important heterogeneity in terms of size and shape and were larger than spheroids generated with the other methods. Spheroids generated in hanging drops, in the Sphericalplate 5D™, and in agarose microwell plates were homogeneous, with well-defined round shape and a mean diameter of 90 µm. GSIS results showed improved insulin secretion in response to glucose in comparison with native islets and self-aggregated spheroids. Spheroids can be generated using different techniques and each of them present advantages and inconveniences. For islet cell aggregation, we recommend, based on our results, to use the hanging drop technique, the agarose microwell plates, or the Sphericalplate 5D™ depending on the experiments, the latter being the only option available for large-scale spheroids production. SAGE Publications 2020-08-04 /pmc/articles/PMC7563811/ /pubmed/32749168 http://dx.doi.org/10.1177/0963689720937292 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Article
Wassmer, Charles-Henri
Bellofatto, Kevin
Perez, Lisa
Lavallard, Vanessa
Cottet-Dumoulin, David
Ljubicic, Sanda
Parnaud, Géraldine
Bosco, Domenico
Berishvili, Ekaterine
Lebreton, Fanny
Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title_full Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title_fullStr Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title_full_unstemmed Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title_short Engineering of Primary Pancreatic Islet Cell Spheroids for Three-dimensional Culture or Transplantation: A Methodological Comparative Study
title_sort engineering of primary pancreatic islet cell spheroids for three-dimensional culture or transplantation: a methodological comparative study
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7563811/
https://www.ncbi.nlm.nih.gov/pubmed/32749168
http://dx.doi.org/10.1177/0963689720937292
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