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Assembly of Human Umbilical Vein Endothelial Cells on Compliant Hydrogels.

ABSTRACT: Angiogenesis is the process by which endothelial cells grow and disassemble into functional blood vessels. In this study, we examine the fundamental processes that control the assembly of endothelial cells into networks in vitro. Network assembly is known to be influenced by matrix mechanics and chemical signals. However, the roles of substrate stiffness and chemical signals in network formation is unclear. In this study, human umbilical vein endothelial cells (HUVECs) were seeded onto RGD or GFOGER functionalized polyacrylamide gels of varying stiffness. Cells were either treated with bFGF, VEGF, or left untreated and observed over time. We found that cells form stable networks on soft gels (Young's modulus 140 Pa) when untreated but that growth factors induce increased cell migration which leads to network instability. On stiffer substrates (Young's modulus 2500 Pa) cells do not assemble into networks either with or without growth factors in any combination. Our results indicate that cells assemble to networks below a critical compliance, that a critical cell density is needed for network formation, and that growth factors can inhibit network formation through an increase in motility.

SUBMITTER: Saunders RL

PROVIDER: S-EPMC3132815 | biostudies-literature | 2010 Mar

REPOSITORIES: biostudies-literature

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Assembly of Human Umbilical Vein Endothelial Cells on Compliant Hydrogels.

Saunders Randi L RL Hammer Daniel A DA

Cellular and molecular bioengineering 20100301 1

Angiogenesis is the process by which endothelial cells grow and disassemble into functional blood vessels. In this study, we examine the fundamental processes that control the assembly of endothelial cells into networks in vitro. Network assembly is known to be influenced by matrix mechanics and chemical signals. However, the roles of substrate stiffness and chemical signals in network formation is unclear. In this study, human umbilical vein endothelial cells (HUVECs) were seeded onto RGD or GF ...[more]

PMID: 21754971

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Project description:β-Cell functionality and survival are highly dependent on the cells' microenvironment and cell-cell interactions. Since the pancreas is a highly vascularized organ, the crosstalk between β-cells and endothelial cells (ECs) is vital to ensure proper function. To understand the interaction of pancreatic β-cells with vascular ECs, we sought to investigate the impact of the spatial distribution on the interaction of human cell line-based β-cells (EndoC-βH3) and human umbilical vein endothelial cells (HUVECs). We focused on the evaluation of three major spatial distributions, which can be found within human islets in vivo, in tissue-engineered heterotypic cell spheroids, so-called pseudo-islets, by controlling the aggregation process using magnetic levitation. We report that heterotypic spheroids formed by spontaneous aggregation cannot be maintained in culture due to HUVEC disassembly over time. In contrast, magnetic levitation allows the formation of stable heterotypic spheroids with defined spatial distribution and significantly facilitated HUVEC integration. To the best of our knowledge, this is the first study that introduces a human-only cell line-based in vitro test system composed of a coculture of β-cells and ECs with a successful stimulation of β-cell secretory function monitored by a glucose-stimulated insulin secretion assays. In addition, we systematically investigate the impact of the spatial distribution on cocultures of human β-cells and ECs, showing that the architecture of pseudo-islets significantly affects β-cell functionality. Impact statement Tissue engineering of coculture systems containing β-cells and endothelial cells (ECs) is a promising technique to stimulate β-cell functionality. In this study, we analyzed human pancreatic islet tissue and revealed three different native distributions of β-cells and ECs. We successfully recreated these distributions in vitro by employing magnetic levitation of human β-cells and ECs, forming controlled heterotypic pseudo-islets, which enabled us to identify a significant impact of the pseudo-islet architecture on insulin secretion.

Dataset Information

Assembly of Human Umbilical Vein Endothelial Cells on Compliant Hydrogels.

Publications

Assembly of Human Umbilical Vein Endothelial Cells on Compliant Hydrogels.

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