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Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

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Analyzing the Relationship between Solid-Phase Molecular Presentation and Cell Proliferation, Morphology and

Enrique Azuaje-Hualde¹, Naiara Lartitegui-Meneses¹, Juncal Alonso-Cabrera^1,2

¹Microfluidics Cluster UPV/EHU, BIOMICs Microfluidics Group, University of the Basque Country UPV/EHU, Vitoria-Gasteiz 01006, Spain.

ACS Applied Materials & Interfaces

|December 29, 2025

Summary

This summary is machine-generated.

This study shows how fibroblast growth factor 2 (FGF2) presentation affects stem cell behavior. CellStudio platform precisely controls FGF2 levels, revealing dose-dependent impacts on cell growth, survival, and secretion.

Keywords:

biosensing cell microenvironment growth factor presentation mesenchymal stem cells secretomics solid-phase

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Area of Science:

Biotechnology
Cell Biology
Biomaterials

Background:

Precise spatial control of molecular cues is crucial for engineering functional cell microenvironments.
CellStudio, a bead-based microsystem, enables localized biosensing and cell analysis with spatial resolution.

Purpose of the Study:

To investigate the influence of solid-phase fibroblast growth factor 2 (FGF2) presentation on mesenchymal stem cell (MSC) behavior.
To extend CellStudio's capabilities for analyzing cellular responses to controlled growth factor stimulation.

Main Methods:

Utilized CellStudio, a bead-based microsystem, for precise spatial presentation of FGF2-functionalized microbeads.
Customized the ratio of FGF2-functionalized microbeads surrounding patterned MSC clusters.
Monitored cell proliferation, survival, morphology, cytoskeletal remodeling, and receptor expression.
Quantified vascular endothelial growth factor (VEGF) secretion using integrated bead sensors.

Main Results:

Demonstrated dose-dependent effects of solid-phase FGF2 on MSC proliferation, survival, and morphology.
Observed FGF2-induced cytoskeletal remodeling, increased cell spreading and elongation.
Showed early upregulation of fibroblast growth factor receptor 1 (FGFR1) membrane expression.
Confirmed enhanced VEGF secretion under local FGF2 stimulation, captured in situ.

Conclusions:

CellStudio provides a versatile platform for high-resolution, parallel analysis of cellular responses to growth factor presentation.
This approach enables detailed investigation of cell-microenvironment interactions.
Potential applications include regenerative medicine, drug screening, and fundamental cell biology research.