BMDCs were embedded into bovine collagen type I matrix (1.6 mg/ml; Advanced BioMatrix) and introduced to a μ-slide chemotaxis chamber (Ibidi) according to the manufacturer’s instructions. Various concentrations of the chemokine CCL19 (Peprotech) in 2% FCS/RPMI were added to the chamber upon solidification of the collagen matrix (37°C for 45 min). Live-cell imaging (5% CO2, 37°C) was performed immediately using a Nikon microscope. Time-lapse images (2- to 3-min intervals between images) were acquired using Metamorph (Molecular Devices). Captured images were processed using the chemotaxis plugin (ImageJ/Fiji).
Bovine collagen type 1 matrix
Manufactured by Advanced BioMatrix
Bovine collagen type I matrix is a purified and lyophilized bovine collagen material. It provides a natural three-dimensional scaffold for cell cultures and tissue engineering applications.
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Lab products found in correlation
2 protocols using bovine collagen type 1 matrix
1
Chemotaxis Assay of Dendritic Cells
2
Collagen-Embedded Spheroid Assay
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Cells were cultured for 72 h as described above to obtain compact spheroids. The latter were embedded into 1.6 mg/mL of bovine collagen type I matrix (Advanced Biomatrix), diluted in 0.12% methylcellulose, and covered with serum-free medium in the presence or absence of inhibitors. Spheroids were then incubated for 24 h in a temperature- and CO2-controlled chamber. Images were taken using an Olympus microscope (Objective 4X). The size of the spheroids was quantified by measuring the area occupied by cells using ImageJ software (rsb.info.nih.gov/ij/ ). Each tested condition was done in sextuplet repeated 3 times.
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