Real-Time Spatiotemporal Measurement of Extracellular Signaling Molecules Using an Aptamer Switch-Conjugated Hydrogel Matrix.

Autor: Park CH; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA.; Department of Radiology, Stanford University, Stanford, CA, 94305, USA.; Department of Chemical and Biological Engineering, Gachon University, Seongnam, 13120, Republic of Korea., Thompson IAP; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA.; Department of Radiology, Stanford University, Stanford, CA, 94305, USA., Newman SS; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Hein LA; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA., Lian X; Department of Radiology, Stanford University, Stanford, CA, 94305, USA., Fu KX; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA.; Department of Radiology, Stanford University, Stanford, CA, 94305, USA.; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, 46556, USA., Pan J; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA.; Department of Radiology, Stanford University, Stanford, CA, 94305, USA.; Department of Mechanical and Aerospace Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, 32611, USA., Eisenstein M; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA., Soh HT; Department of Electrical Engineering, Stanford University, Stanford, CA, 94305, USA.; Department of Radiology, Stanford University, Stanford, CA, 94305, USA.
Jazyk: angličtina
Zdroj: Advanced materials (Deerfield Beach, Fla.) [Adv Mater] 2024 Jan; Vol. 36 (4), pp. e2306704. Date of Electronic Publication: 2023 Nov 29.
DOI: 10.1002/adma.202306704
Abstrakt: Cells rely on secreted signaling molecules to coordinate essential biological functions including development, metabolism, and immunity. Unfortunately, such signaling processes remain difficult to measure with sufficient chemical specificity and temporal resolution. To address this need, an aptamer-conjugated hydrogel matrix that enables continuous fluorescent measurement of specific secreted analytes - in two dimensions, in real-time is developed. As a proof of concept, real-time imaging of inter-cellular cyclic adenosine 3',5'-monophosphate (cAMP) signals in Dictyostelium discoideum amoeba cells is performed. A set of aptamer switches that generate a rapid and reversible change in fluorescence in response to cAMP signals is engineered. By combining multiple switches with different dynamic ranges, measure cAMP concentrations spanning three orders of magnitude in a single experiment can be measured. These sensors are embedded within a biocompatible hydrogel on which cells are cultured and their cAMP secretions can be imaged using fluorescent microscopy. Using this aptamer-hydrogel material system, the first direct measurements of oscillatory cAMP signaling that correlate closely with previous indirect measurements are achieved. Using different aptamer switches, this approach can be generalized for measuring other secreted molecules to directly visualize diverse extracellular signaling processes and the biological effects that they trigger in recipient cells.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
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