Abstract
Recent studies using in vivo models have characterized lymph flow and demonstrated that lymph flow plays a key role in the later stages of lymphatic vascular development, including vascular remodeling, to create a hierarchical collecting vessel network and lymphatic valves (Sweet et al., J Clin Invest 125, 2995–3007, 2015). However, mechanistic insights into the response of lymphatic endothelial cells to fluid flow are difficult to obtain from in vivo studies because of the small size of lymphatic vessels and the technical challenge of lymphatic endothelial cell isolation. On the other hand, in vitro experiments can be tailored to isolate and test specific mechanotransduction pathways more cleanly than conditions in vivo. To measure in vitro the cellular response to flow, cultured primary lymphatic endothelial cells can be exposed to highly specific fluid forces like those believed to exist in vivo. Such in vitro studies have recently helped identify FOXC2 and GATA2 as important transcriptional regulators of lymphatic function during valve formation that are regulated by lymph flow dynamics. This chapter discusses the methods used to expose primary lymphatic endothelial cells (LECs) to lymph fluid dynamics and the relationship of these in vitro studies to in vivo lymphatic biology.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sweet DT et al (2015) Lymph flow regulates collecting lymphatic vessel maturation in vivo. J Clin Invest 125:2995–3007
Davies PF (1995) Flow-mediated endothelial mechanotransduction. Physiol Rev 75:519–560
DePaola N et al (1999) Spatial and temporal regulation of gap junction connexin43 in vascular endothelial cells exposed to controlled disturbed flows in vitro. Proc Natl Acad Sci 96:3154–3159
Passerini AG et al (2005) Regional determinants of arterial endothelial phenotype dominate the impact of gender or short-term exposure to a high-fat diet. Biochem Biophys Res Commun 332:142–148
Sabine A et al (2015) FOXC2 and fluid shear stress stabilize postnatal lymphatic vasculature. J Clin Invest 125:3861–3877
Kazenwadel J et al (2015) GATA2 is required for lymphatic vessel valve development and maintenance. J Clin Invest 125:2879–2994
Sabine A et al (2012) Mechanotransduction, PROX1, and FOXC2 Cooperate to Control Connexin37 and Calcineurin during Lymphatic-Valve Formation. Dev Cell 22:430–445
Levesque MJ, Nerem RM (1985) The elongation and orientation of cultured endothelial cells in response to shear stress. J Biomech Eng 107:341–347
Davies PF, Remuzzi A, Gordon EJ, Dewey CF, Gimbrone MA (1986) Turbulent fluid shear stress induces vascular endothelial cell turnover in vitro. Proc Natl Acad Sci U S A 83:2114–2117
Chin LK et al (2011) Production of reactive oxygen species in endothelial cells under different pulsatile shear stresses and glucose concentrations. Lab Chip 11:1856
Helmke BP, Davies PF (2002) The cytoskeleton under external fluid mechanical forces: Hemodynamic forces acting on the endothelium. Ann Biomed Eng 30:284–296
Kuo YC et al (2015) Oscillatory shear stress mediates directional reorganization of actin cytoskeleton and alters differentiation propensity of mesenchymal stem cells. Stem Cells 33:429–442
Estrada R et al (2011) Endothelial cell culture model for replication of physiological profiles of pressure, flow, stretch, and shear stress in vitro. Anal Chem 83:3170–3177
Vogel M, Franke J, Frank W, Schroten H (2007) Flow in the well: computational fluid dynamics is essential in flow chamber construction. Cytotechnology 55:41–54
Jiménez JM et al (2014) Macro- and microscale variables regulate stent haemodynamics, fibrin deposition and thrombomodulin expression. J R Soc Interface 11:20131079
Zhang ZL, Crozatier C, Le Berre M, Chen Y (2005) In situ bio-functionalization and cell adhesion in microfluidic devices. Microelectron Eng 78–79:556–562
Choi I et al (2011) Visualization of lymphatic vessels by Prox1-promoter directed GFP reporter in a bacterial artificial chromosome-based transgenic mouse. Blood 117:362–365
Deng Y, Atri D, Eichmann A, Simons M (2013) Endothelial ERK signaling controls lymphatic fate specification. J Clin Invest 123:1202–1215
Levet S et al (2013) Bone morphogenetic protein 9 (BMP9) controls lymphatic vessel maturation and valve formation. Blood 122:598–607
Shimahara A, Yamakawa N, Nishikata I, Morishita K (2010) Acetylation of lysine 564 adjacent to the C-terminal binding protein-binding motif in EVI1 is crucial for transcriptional activation of GATA2. J Biol Chem 285:16967–16977
Ahn EE et al (2013) SON protein regulates GATA-2 through transcriptional control of the MicroRNA 23a 27a 24-2 cluster. J Biol Chem 288:5381–5388
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Sweet, D.T., Hall, J.D., Welsh, J., Kahn, M.L., Jiménez, J.M. (2018). Investigating Effects of Fluid Shear Stress on Lymphatic Endothelial Cells. In: Oliver, G., Kahn, M. (eds) Lymphangiogenesis. Methods in Molecular Biology, vol 1846. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8712-2_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8712-2_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8711-5
Online ISBN: 978-1-4939-8712-2
eBook Packages: Springer Protocols