Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells

V Sordi, A Ferri, V Ceserani, E Ciusani, E Dugnani, Silvia Pellegrini, R Nano, L Pecciarini, Augusto Pessina, L Pascucci, L Piemonti, G Alessandri

Research output: Contribution to journalArticle

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Abstract

BACKGROUND: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. METHODS: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics. RESULTS: EC were obtained from 25% of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC). DISCUSSION: This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)141-152
Number of pages12
JournalCytotherapy
Volume19
Issue number1
DOIs
Publication statusPublished - 2017

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Islets of Langerhans
Endothelial Cells
Islets of Langerhans Transplantation
Regenerative Medicine
Vascular Endothelial Growth Factor Receptor-1
Cell Line
Chemokine CCL2
von Willebrand Factor
Transmission Electron Microscopy
Interleukin-8
Epidermal Growth Factor
Protein-Tyrosine Kinases
Vascular Endothelial Growth Factor A
Endothelium
Fluorescent Antibody Technique
Pancreas
Melanoma
Flow Cytometry
Cell Culture Techniques
Phenotype

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Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells. / Sordi, V; Ferri, A; Ceserani, V; Ciusani, E; Dugnani, E; Pellegrini, Silvia; Nano, R; Pecciarini, L; Pessina, Augusto; Pascucci, L; Piemonti, L; Alessandri, G.

In: Cytotherapy, Vol. 19, No. 1, 2017, p. 141-152.

Research output: Contribution to journalArticle

Sordi, V ; Ferri, A ; Ceserani, V ; Ciusani, E ; Dugnani, E ; Pellegrini, Silvia ; Nano, R ; Pecciarini, L ; Pessina, Augusto ; Pascucci, L ; Piemonti, L ; Alessandri, G. / Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells. In: Cytotherapy. 2017 ; Vol. 19, No. 1. pp. 141-152.
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abstract = "BACKGROUND: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. METHODS: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics. RESULTS: EC were obtained from 25{\%} of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC). DISCUSSION: This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes. Copyright {\circledC} 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.",
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T1 - Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells

AU - Sordi, V

AU - Ferri, A

AU - Ceserani, V

AU - Ciusani, E

AU - Dugnani, E

AU - Pellegrini, Silvia

AU - Nano, R

AU - Pecciarini, L

AU - Pessina, Augusto

AU - Pascucci, L

AU - Piemonti, L

AU - Alessandri, G

PY - 2017

Y1 - 2017

N2 - BACKGROUND: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. METHODS: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics. RESULTS: EC were obtained from 25% of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC). DISCUSSION: This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

AB - BACKGROUND: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. METHODS: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics. RESULTS: EC were obtained from 25% of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC). DISCUSSION: This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

U2 - 10.1016/j.jcyt.2016.10.005

DO - 10.1016/j.jcyt.2016.10.005

M3 - Article

VL - 19

SP - 141

EP - 152

JO - Cytotherapy

JF - Cytotherapy

SN - 1465-3249

IS - 1

ER -