Effect of Collagen-Coating Variations on the Morphology and Viability of Human Vascular Smooth Muscle Cells

Abstract

Purpose: Collagen is a critical extracellular matrix (ECM) component that significantly influences cellular behaviors such as adhesion, migration, and proliferation. Optimizing collagen coating protocols is essential for developing accurate in vitro models, particularly for studying vascular smooth muscle cells (HVSMCs). The aim of this study was to optimize collagen coating protocols for in vitro models using HVSMCs by assessing cell morphology, adhesion potential, and viability under various collagen concentrations and incubation conditions. Methods: HVSMCs were cultured on surfaces coated with different concentrations of Type 1 Rat Tail Collagen with different cell number (as 104 cells/well and 204 cells/well). The cells were incubated at various temperatures (4°C, 25°C, and 37°C). Morphological analysis was performed using phase-contrast microscopy to observe the alignment and phenotype of the cells. Cell adhesion was assessed using DAPI staining, and cell viability was evaluated using the Presto Blue assay after 96 hours of incubation. Results: Collagen coating significantly influenced HVSMC behavior. The cells transitioned to a contractile phenotype, evidenced by tight, parallel bundle alignment, which is critical for maintaining vascular tone. Enhanced cell adhesion was observed in specific collagen-coated groups across different temperatures, particularly in the F, G, and H groups. Additionally, collagen coating did not significantly increase cell proliferation, making it suitable for in vitro vascular models. Optimal results were observed in groups seeded with 104 cells and incubated at 25°C and 37°C. Conclusion: The study highlights the importance of optimizing extracellular matrix components like collagen in developing functional in vitro models. The identified optimal conditions for collagen coating will be valuable for future vascular modeling studies, providing a reliable foundation for in vitro research.

Keywords

• Collagen-coating
• in vitro vascular model
• human vascular smooth muscle cells
• morphology
• viability

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