Introduction Monocyte chemoattractant protein-1 (MCP-1) is a bioactive molecule that is expressed in significant amounts in all stages of atherosclerosis. The role of MCP-1 in this disease is to recruit monocytes across the endothelium and into the arterial tissue. Eventually, the monocytes differentiate into cholesterol-engorged macrophages called “foam cells” that result in atherosclerotic plaque formation. The mechanism that MCP-1 uses to mediate monocyte transendothelial migration is believed to be via its concentration gradient. However, the formation of the MCP-1 concentration gradient in the extracellular matrix is still poorly understood. Methods A 3D in vitro vascular tissue model has been developed to study the cellular mechanisms involved in the early stages of atherosclerosis. In the present study, a mathematical model is used to determine the gradient of MCP-1 in the collagen matrix of the 3D in vitro vascular tissue model. Experiments were performed to investigate the stability of MCP-1 and the interaction between MCP-1 and the collagen matrix. Results and Conclusions MCP-1 is stable for at least 24 hours under experimental conditions and MCP-1 interacts with the collagen matrix. The diffusion coefficient for the transport of MCP-1 in the collagen matrix and the rate constant for the binding of MCP-1 to collagen were determined to be 0.108 mm2 hr−1 and 0.858 hr−1, respectively. Numerical results from the model indicate that the concentration gradients of both soluble and matrix-bound (or static) MCP-1 are formed inside the collagen matrix.
Monocyte chemoattractant protein-1 (MCP-1) is commercially available in a form of recombinant protein. This makes it more convenient to study the functions of MCP-1 and its involvement in many cell functions. However, when using MCP-1 in experimental studies, if the analysis is not performed immediately, the stability of recombinant MCP-1 may become an issue. In this study, the stability of recombinant MCP-1 at different concentrations and storage conditions was investigated. Results show that no significant loss of MCP-1 is observed when MCP-1 solutions were stored at non-freezing condition (4°C) for seven days. However, for storage at freezing conditions (−20°C or −81°C), it appears that the first freeze-thaw cycle may contribute to some loss of MCP-1 in the solutions, and such loss may be concentration and time dependent. The effect of multiple freeze-thaw cycles for storage at freezing conditions was also examined. Data reveal that the second freeze-thaw cycle causes approximately 50% loss of MCP-1 in the solutions. This finding confirms that multiple freeze-thaw cycles should be avoided. The findings of this study provide an outline of how storage can affect the stability of recombinant proteins and should be taken into account during the evaluation of the concentration of recombinant proteins.
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