Sprouting angiogenesis under a chemical gradient regulated by interactions with an endothelial monolayer in a microfluidic platform.

Microfluidic cell culture assays are versatile tools for studying cell migration, particularly angiogenesis. Such assays can deliver precisely controlled linear gradients of chemical stimuli to cultured cells in a microfluidic channel, offering excellent optical resolution and in situ monitoring of cellular morphogenesis in response to a gradient. Microfluidic cell culture assays provide a chemical gradient subject to molecular diffusion, although cellular metabolism can perturb it. The actual gradient perturbed by cells has not been precisely described in the context of regulated cellular morphogenesis. We modeled the chemical gradient in a microfluidic channel by simulating the analyte(VEGF) distribution during cellular interactions. The results were experimentally verified by monitoring sprouting angiogenic response from a monolayer of human umbilical vein endothelial cells (hUVECs) into a type 1 collagen scaffold. The simulation provided a basis for understanding a real distribution of the analyte interrupted by cells in microfluidic device. The new protocol enables one to quantify the morphogenesis of hUVECs under a flat, less-steep, or steep gradient.

[1]  John A. Pedersen,et al.  Mechanobiology in the Third Dimension , 2005, Annals of Biomedical Engineering.

[2]  L. Griffith,et al.  Capturing complex 3D tissue physiology in vitro , 2006, Nature Reviews Molecular Cell Biology.

[3]  Roger D Kamm,et al.  Biomechanical Regulation of Endothelium-dependent Events Critical for Adaptive Remodeling* , 2009, Journal of Biological Chemistry.

[4]  Craig A Simmons,et al.  Macro- and microscale fluid flow systems for endothelial cell biology. , 2010, Lab on a chip.

[5]  Kenneth M. Yamada,et al.  Modeling Tissue Morphogenesis and Cancer in 3D , 2007, Cell.

[6]  Tharathorn Rimchala,et al.  Surface‐Treatment‐Induced Three‐Dimensional Capillary Morphogenesis in a Microfluidic Platform , 2009, Advanced materials.

[7]  S. Thorslund,et al.  A fluidic device to study directional angiogenesis in complex tissue and organ culture models. , 2009, Lab on a chip.

[8]  Janet Rossant,et al.  Endothelial cells and VEGF in vascular development , 2005, Nature.

[9]  L. Griffith,et al.  Transport‐mediated angiogenesis in 3D epithelial coculture , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  Vernella Vickerman,et al.  Design, fabrication and implementation of a novel multi-parameter control microfluidic platform for three-dimensional cell culture and real-time imaging. , 2008, Lab on a chip.

[11]  T W Gardner,et al.  Effect of vascular endothelial growth factor on cultured endothelial cell monolayer transport properties. , 2000, Microvascular research.

[12]  Federica Boschetti,et al.  Synergy between interstitial flow and VEGF directs capillary morphogenesis in vitro through a gradient amplification mechanism. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Raquel Perez-Castillejos,et al.  Partitioning microfluidic channels with hydrogel to construct tunable 3-D cellular microenvironments. , 2008, Biomaterials.

[14]  William J. Polacheck,et al.  Interstitial flow influences direction of tumor cell migration through competing mechanisms , 2011, Proceedings of the National Academy of Sciences.

[15]  Hanry Yu,et al.  A novel 3D mammalian cell perfusion-culture system in microfluidic channels. , 2007, Lab on a chip.

[16]  R. Auerbach,et al.  Assays for angiogenesis: a review. , 1991, Pharmacology & therapeutics.

[17]  Kenneth M. Yamada,et al.  Cell migration in 3D matrix. , 2005, Current opinion in cell biology.

[18]  R. Kamm,et al.  The Stiffness of Three-dimensional Ionic Self-assembling Peptide Gels Affects the Extent of Capillary-like Network Formation , 2007, Cell Biochemistry and Biophysics.

[19]  Judah Folkman,et al.  Angiogenesis in vitro , 1980, Nature.

[20]  Minseok S. Kim,et al.  A microfluidic platform for 3-dimensional cell culture and cell-based assays , 2007, Biomedical microdevices.

[21]  Michela Matteoli,et al.  Overflow microfluidic networks for open and closed cell cultures on chip. , 2010, Analytical chemistry.

[22]  A. Lee,et al.  Engineering microscale cellular niches for three-dimensional multicellular co-cultures. , 2009, Lab on a chip.

[23]  Andreas Manz,et al.  Latest developments in microfluidic cell biology and analysis systems. , 2010, Analytical chemistry.

[24]  Hanry Yu,et al.  A gel-free 3D microfluidic cell culture system. , 2008, Biomaterials.

[25]  R. Kamm,et al.  Cell migration into scaffolds under co-culture conditions in a microfluidic platform. , 2009, Lab on a chip.

[26]  Bingcheng Lin,et al.  A microfluidic device for characterizing the invasion of cancer cells in 3‐D matrix , 2009, Electrophoresis.

[27]  Robert Langer,et al.  EDITORIAL: TISSUE ENGINEERING: PERSPECTIVES, CHALLENGES, AND FUTURE DIRECTIONS , 2007 .

[28]  O. Hudlická,et al.  What makes blood vessels grow? , 1991, The Journal of physiology.

[29]  Göran Stemme,et al.  A microfluidic device for parallel 3‐D cell cultures in asymmetric environments , 2007, Electrophoresis.

[30]  Gi Seok Jeong,et al.  Microfluidic assay of endothelial cell migration in 3D interpenetrating polymer semi-network HA-Collagen hydrogel , 2011, Biomedical microdevices.

[31]  J M Piret,et al.  Cytokine manipulation of primitive human hematopoietic cell self-renewal. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Yongdoo Park,et al.  Synthesis of cell-laden alginate hollow fibers using microfluidic chips and microvascularized tissue-engineering applications. , 2009, Small.

[33]  Roger D. Kamm,et al.  Microfluidic Platforms for Studies of Angiogenesis, Cell Migration, and Cell–Cell Interactions , 2010, Annals of Biomedical Engineering.

[34]  Napoleone Ferrara,et al.  Angiogenesis as a therapeutic target , 2005, Nature.

[35]  B. Chung,et al.  Generation of stable concentration gradients in 2D and 3D environments using a microfluidic ladder chamber , 2007, Biomedical microdevices.

[36]  R. Kamm,et al.  Concentration gradients in microfluidic 3D matrix cell culture systems , 2010 .

[37]  Rakesh K. Jain,et al.  Quantitative angiogenesis assays: Progress and problems , 1997, Nature Medicine.

[38]  Shuichi Takayama,et al.  Regulating microenvironmental stimuli for stem cells and cancer cells using microsystems. , 2010, Integrative biology : quantitative biosciences from nano to macro.