Localized Cell-Surface Sampling of a Secreted Factor Using Cell-Targeting Beads.

Anal Chem

Department of Chemistry, University of Washington, Box 351700, Seattle, Washington 98195, United States.

Published: October 2020

Intercellular communication through the secretion of soluble factors plays a vital role in a wide range of biological processes (e.g., homeostasis, immune response), yet identification and quantification of many of these factors can be challenging due to their degradation or sequestration in cell culture media prior to analysis. Here, we present a customizable bead-based system capable of simultaneously binding to live cells (through antibody-mediated cell tethering) and capturing cell-secreted molecules. Our functionalized beads capture secreted molecules (e.g., hepatocyte growth factor secreted by fibroblasts) that are diminished when sampled via traditional supernatant analysis techniques ( < 0.05), effectively rescuing a reduced signal in the presence of neutralizing components in the cell culture media. Our system enables capture and analysis of molecules integral to chemical communication that would otherwise be markedly decreased prior to analysis.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8259888PMC
http://dx.doi.org/10.1021/acs.analchem.0c02578DOI Listing

Publication Analysis

Top Keywords

cell culture
8
culture media
8
prior analysis
8
localized cell-surface
4
cell-surface sampling
4
sampling secreted
4
secreted factor
4
factor cell-targeting
4
cell-targeting beads
4
beads intercellular
4

Similar Publications

Background: Sepsis, a critical global health challenge, accounted for approximately 20% of worldwide deaths in 2017. Although the Sequential Organ Failure Assessment (SOFA) score standardizes the diagnosis of organ dysfunction, early sepsis detection remains challenging due to its insidious symptoms. Current diagnostic methods, including clinical assessments and laboratory tests, frequently lack the speed and specificity needed for timely intervention, particularly in vulnerable populations such as older adults, intensive care unit (ICU) patients, and those with compromised immune systems.

View Article and Find Full Text PDF

Immunologic bile duct destruction is a pathogenic condition associated with vanishing bile duct syndrome (VBDS) after liver transplantation and hematopoietic stem-cell transplantation. As the bile acid receptor sphingosine 1-phosphate receptor 2 (S1PR2) plays a critical role in recruitment of bone marrow-derived monocytes/macrophages to sites of cholestatic liver injury, S1PR2 expression was examined using cultured macrophages and patient tissues. Bile canaliculi destruction precedes intrahepatic ductopenia; therefore, we focused on hepatocyte S1PR2 and the downstream RhoA/Rho kinase 1 (ROCK1) signaling pathway and bile canaliculi alterations using three-dimensional hepatocyte culture models that form obvious bile canaliculus-like networks.

View Article and Find Full Text PDF

Variants in the SOX9 transactivation middle domain induce axial skeleton dysplasia and scoliosis.

Proc Natl Acad Sci U S A

January 2025

Department of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China.

SOX9 is a crucial transcriptional regulator of cartilage development and homeostasis. Dysregulation of is associated with a wide spectrum of skeletal disorders, including campomelic dysplasia, acampomelic campomelic dysplasia, and scoliosis. Yet how variants contribute to the spectrum of axial skeletal disorders is not well understood.

View Article and Find Full Text PDF

Purpose: A human model able to simulate the manifestation of corneal endothelium decompensation could be advantageous for wound healing and future cell therapy assessment. The study aimed to establish an ex vivo human cornea endothelium wound model where endothelium function can be evaluated by measuring corneal thickness changes.

Methods: The human cornea was maintained in an artificial anterior chamber, with a continuous culture medium infusion system designed to sustain corneal endothelium and epithelium simultaneously.

View Article and Find Full Text PDF

Tendon injuries and disorders associated with mechanical tendon overuse are common musculoskeletal problems. Even though tendons play a central role in human movement, the intrinsic healing process of tendon is very slow. So far, it is known that tendon cell activity is supported by several interstitial cells within the tendon.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!