Protocol for in vivo and in vitro study of lymphatic valve formation driven by shear stress signaling pathway

Richa Banerjee; Luz A Knauer; Ying Yang

doi:10.1016/j.xpro.2023.102141

Protocol for in vivo and in vitro study of lymphatic valve formation driven by shear stress signaling pathway

STAR Protoc. 2023 Apr 17;4(2):102141. doi: 10.1016/j.xpro.2023.102141. Online ahead of print.

Authors

Richa Banerjee¹, Luz A Knauer¹, Ying Yang²

Affiliations

¹ Department of Molecular Pharmacology & Physiology, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA.
² Department of Molecular Pharmacology & Physiology, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA. Electronic address: yingyang@usf.edu.

Abstract

Here we detail a protocol for the isolation and processing of lymphatic enriched tissue of mouse models for the purpose of immunostaining and quantification of lymphatic valves, vessel length, and vessel diameter. Furthermore, we describe an optimized protocol for exposing treated human dermal lymphatic endothelial cells to flow for the purpose of studying lymph shear stress responses via gene expression and protein detection methods. This approach is useful to study lymphatic valve formation driven by oscillatory shear stress. For complete details on the use and execution of this protocol, please refer to Scallan et al. (2021).¹.

Keywords: Cell culture; Developmental biology; Gene expression; Microscopy; Model organisms; Molecular biology.

Grants and funding

R01 HL145397/HL/NHLBI NIH HHS/United States