Fibronectin and Collagen IV Microcontact Printing Improves Insulin Secretion by INS1E Cells

Elahe Hadavi, Jeroen Leijten, Jenny Brinkmann, Pascal Jonkheijm, Marcel Karperien, Aart van Apeldoorn (Corresponding Author)

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Abstract

Extracellular matrix (ECM) molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation, and their endocrine function. During clinical islet transplantation, the β-cell's ECM environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin (FN), collagen type IV (Col4), and laminin (LN), are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells. It is also thought that three-dimensional (3D) culture of β-cells can improve glucose responsiveness by providing a specific niche, in which cells can interact with each other in a more natural manner. Conventional suspension cultures with β-cells results generally in a heterogeneous population with small and large aggregates, in which cells experience different nutrient diffusion limitations, negatively affecting their physiology and function. In this study, we have explored the effect of FN, Col4, and LN111 on INS1E insulinoma cells by using microcontact printing (μCP) to investigate whether a controlled environment and aggregate dimensions would improve their endocrine function. Using this method, we produced a pattern of well-defined circular spots of FN, Col4, and LN111 on polydimethylsiloxane with high spatial resolution. Cell seeding of the INS1E cells on these ECM protein spots resulted in the formation of 3D β-cell aggregates. We show that these INS1E aggregates have very reproducible dimensions, and that the cell culture method can be easily adjusted, leading to a highly accurate way of forming 3D β-cell aggregates on an ECM-functionalized substrate. In addition, we show that ECM molecules can act as anchoring points for β-cells on an otherwise non-cell-adherent material, and this can improve both the endocrine function and viability. We found a significant increase in the secretion of insulin by INS1E cells cultured on μCP FN and Col4 substrates, in comparison to cells that were growing in monolayers on substrates without ECM molecules. Moreover, INS1E cells growing on circular ECM spots in a 3D manner showed improved endocrine function in comparison to their two-dimensional counterparts. This research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.

Original languageEnglish
Pages (from-to)628-636
Number of pages9
JournalTissue engineering. Part C, Methods
Volume24
Issue number11
DOIs
Publication statusPublished - 15 Nov 2018

Fingerprint

Printing
Insulin
Fibronectins
Collagen
Extracellular Matrix
Molecules
Cell Culture Techniques
Substrates
Islets of Langerhans
Cell Survival
Collagen Type IV
Extracellular Matrix Proteins
Physiology
Cell proliferation
Laminin
Polydimethylsiloxane
Medical problems
Cell culture
Cellular Microenvironment
Controlled Environment

Keywords

  • islet transplantation
  • type 1 diabetes
  • β-cell replacement therapies
  • β-cells
  • insulin secretion

Cite this

@article{96003898f03f4cc9b73ad59cb26a638b,
title = "Fibronectin and Collagen IV Microcontact Printing Improves Insulin Secretion by INS1E Cells",
abstract = "Extracellular matrix (ECM) molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation, and their endocrine function. During clinical islet transplantation, the β-cell's ECM environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin (FN), collagen type IV (Col4), and laminin (LN), are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells. It is also thought that three-dimensional (3D) culture of β-cells can improve glucose responsiveness by providing a specific niche, in which cells can interact with each other in a more natural manner. Conventional suspension cultures with β-cells results generally in a heterogeneous population with small and large aggregates, in which cells experience different nutrient diffusion limitations, negatively affecting their physiology and function. In this study, we have explored the effect of FN, Col4, and LN111 on INS1E insulinoma cells by using microcontact printing (μCP) to investigate whether a controlled environment and aggregate dimensions would improve their endocrine function. Using this method, we produced a pattern of well-defined circular spots of FN, Col4, and LN111 on polydimethylsiloxane with high spatial resolution. Cell seeding of the INS1E cells on these ECM protein spots resulted in the formation of 3D β-cell aggregates. We show that these INS1E aggregates have very reproducible dimensions, and that the cell culture method can be easily adjusted, leading to a highly accurate way of forming 3D β-cell aggregates on an ECM-functionalized substrate. In addition, we show that ECM molecules can act as anchoring points for β-cells on an otherwise non-cell-adherent material, and this can improve both the endocrine function and viability. We found a significant increase in the secretion of insulin by INS1E cells cultured on μCP FN and Col4 substrates, in comparison to cells that were growing in monolayers on substrates without ECM molecules. Moreover, INS1E cells growing on circular ECM spots in a 3D manner showed improved endocrine function in comparison to their two-dimensional counterparts. This research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.",
keywords = "islet transplantation, type 1 diabetes, β-cell replacement therapies, β-cells, insulin secretion",
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Fibronectin and Collagen IV Microcontact Printing Improves Insulin Secretion by INS1E Cells. / Hadavi, Elahe; Leijten, Jeroen; Brinkmann, Jenny; Jonkheijm, Pascal; Karperien, Marcel; van Apeldoorn, Aart (Corresponding Author).

In: Tissue engineering. Part C, Methods, Vol. 24, No. 11, 15.11.2018, p. 628-636.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Fibronectin and Collagen IV Microcontact Printing Improves Insulin Secretion by INS1E Cells

AU - Hadavi, Elahe

AU - Leijten, Jeroen

AU - Brinkmann, Jenny

AU - Jonkheijm, Pascal

AU - Karperien, Marcel

AU - van Apeldoorn, Aart

PY - 2018/11/15

Y1 - 2018/11/15

N2 - Extracellular matrix (ECM) molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation, and their endocrine function. During clinical islet transplantation, the β-cell's ECM environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin (FN), collagen type IV (Col4), and laminin (LN), are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells. It is also thought that three-dimensional (3D) culture of β-cells can improve glucose responsiveness by providing a specific niche, in which cells can interact with each other in a more natural manner. Conventional suspension cultures with β-cells results generally in a heterogeneous population with small and large aggregates, in which cells experience different nutrient diffusion limitations, negatively affecting their physiology and function. In this study, we have explored the effect of FN, Col4, and LN111 on INS1E insulinoma cells by using microcontact printing (μCP) to investigate whether a controlled environment and aggregate dimensions would improve their endocrine function. Using this method, we produced a pattern of well-defined circular spots of FN, Col4, and LN111 on polydimethylsiloxane with high spatial resolution. Cell seeding of the INS1E cells on these ECM protein spots resulted in the formation of 3D β-cell aggregates. We show that these INS1E aggregates have very reproducible dimensions, and that the cell culture method can be easily adjusted, leading to a highly accurate way of forming 3D β-cell aggregates on an ECM-functionalized substrate. In addition, we show that ECM molecules can act as anchoring points for β-cells on an otherwise non-cell-adherent material, and this can improve both the endocrine function and viability. We found a significant increase in the secretion of insulin by INS1E cells cultured on μCP FN and Col4 substrates, in comparison to cells that were growing in monolayers on substrates without ECM molecules. Moreover, INS1E cells growing on circular ECM spots in a 3D manner showed improved endocrine function in comparison to their two-dimensional counterparts. This research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.

AB - Extracellular matrix (ECM) molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation, and their endocrine function. During clinical islet transplantation, the β-cell's ECM environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin (FN), collagen type IV (Col4), and laminin (LN), are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells. It is also thought that three-dimensional (3D) culture of β-cells can improve glucose responsiveness by providing a specific niche, in which cells can interact with each other in a more natural manner. Conventional suspension cultures with β-cells results generally in a heterogeneous population with small and large aggregates, in which cells experience different nutrient diffusion limitations, negatively affecting their physiology and function. In this study, we have explored the effect of FN, Col4, and LN111 on INS1E insulinoma cells by using microcontact printing (μCP) to investigate whether a controlled environment and aggregate dimensions would improve their endocrine function. Using this method, we produced a pattern of well-defined circular spots of FN, Col4, and LN111 on polydimethylsiloxane with high spatial resolution. Cell seeding of the INS1E cells on these ECM protein spots resulted in the formation of 3D β-cell aggregates. We show that these INS1E aggregates have very reproducible dimensions, and that the cell culture method can be easily adjusted, leading to a highly accurate way of forming 3D β-cell aggregates on an ECM-functionalized substrate. In addition, we show that ECM molecules can act as anchoring points for β-cells on an otherwise non-cell-adherent material, and this can improve both the endocrine function and viability. We found a significant increase in the secretion of insulin by INS1E cells cultured on μCP FN and Col4 substrates, in comparison to cells that were growing in monolayers on substrates without ECM molecules. Moreover, INS1E cells growing on circular ECM spots in a 3D manner showed improved endocrine function in comparison to their two-dimensional counterparts. This research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.

KW - islet transplantation

KW - type 1 diabetes

KW - β-cell replacement therapies

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