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In Vitro Microvessel Growth and Remodeling within a Three-Dimensional Microfluidic Environment

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Abstract

This paper presents in vitro microvascular network formation within 3D gel scaffolds made from different concentrations of type-I collagen, fibrin, or a mixture of collagen and fibrin, using a simple microfluidic platform. Initially, microvascular network formation of human umbilical vein endothelial cells was examined using live time-lapse confocal microscopy every 90 min from 3 h to 12 h after seeding within three different concentrations of collagen gel scaffolds. Among the three collagen gel concentrations, the number of skeletons was consistently the highest at 3.0 mg/mL, followed by those of collagen gel scaffolds at 2.5 mg/mL and 2.0 mg/mL. Results demonstrated that concentration of collagen gel scaffolds, which influences matrix stiffness and ligand density, may affect microvascular network formation during the early stages of vasculogenesis. In addition, the maturation of microvascular networks in monoculture under different gel compositions within gel scaffolds (2.5 mg/mL) was examined for 7 days using live confocal microscopy. It was confirmed that pure fibrin gel scaffolds are preferable to collagen gel or collagen/fibrin combinations, significantly reducing matrix retractions during maturation of microvascular networks for 7 days. Finally, early steps in the maturation process of microvascular networks for 14 days were characterized by demonstrating sequential steps of branching, expanding, remodeling, pruning, and clear delineation of lumens within fibrin gel scaffolds. Our findings demonstrate an in vitro model for generating mature microvascular networks within 3D microfluidic fibrin gel scaffolds (2.5 mg/mL), and furthermore suggest the importance of gel concentration and composition in promoting the maturation of microvascular networks.

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Acknowledgments

We would like to thank Lee L. S. Ong and M. S. Chong for very useful discussions. HUVECs were kindly provided by Dr. M. S. Chong (Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore). We also thank Sihua Huang for her support with preparing devices and Grace S. Park for her assistance with figure preparation. This work was supported by funding from the Singapore-MIT Alliance for Research and Technology (SMART) Center.

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Authors and Affiliations

  1. Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology Center, Singapore, 117543, Singapore

    Young K. Park, Ting-Yuan Tu, Sei Hien Lim & Roger D. Kamm

  2. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Young K. Park & Roger D. Kamm

  3. Computational Biology Programme, Department of Biological Sciences, Ntional University of Singapore, Singapore, 119077, Singapore

    Ivan J. M. Clement

  4. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Se Y. Yang & Roger D. Kamm

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  1. Young K. Park
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Correspondence to Roger D. Kamm.

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Associate Editor Philip Leduc oversaw the review of this article.

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Park, Y.K., Tu, TY., Lim, S.H. et al. In Vitro Microvessel Growth and Remodeling within a Three-Dimensional Microfluidic Environment. Cel. Mol. Bioeng. 7, 15–25 (2014). https://doi.org/10.1007/s12195-013-0315-6

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