Collagen matrices were prepared from Nutragen Type I Bovine Collagen (Advanced BioMatrix) in 1X MEM Eagle supplemented with 7.5% sodium bicarbonate (pH 9) in optical-grade black 96-well plates. Matrices were allowed to polymerize at 37°C for 60 min to overnight, and then equilibrated ≥ 60 min in monocyte growth media supplemented with 3% serum. In each well, 5×104 monocytes or macrophages pre-labeled with Cell Trace CFSE (Invitrogen) and pre-infected with T. gondii (THP-1: MOI 4.5 with ME49 mCherry, 4 h; RAW 264.7: MOI 3 with ME49 mCherry) were allowed to migrate into the matrix. After overnight migration at 37°C and 5% CO2, the samples were rinsed with PBS and fixed with 4% formaldehyde. Z-stack image series of the monocytes captured within the collagen matrices were captured with the above-described spinning disk microscopy system and an EC Plan-Neofluar 10x (N.A. 0.30) objective. Volocity software was used to generate 3-D renderings of the Z-stacks and locate each monocyte using an automated Object Finding protocol based on fluorescence intensity and cell size. Vertical distance migrated was calculated as the distance between the centroid of the cell with the highest Z-position and every other cell.
Nutragen type 1 bovine collagen
Manufactured by Advanced BioMatrix
Nutragen Type I Bovine Collagen is a high-quality raw material sourced from bovine connective tissue. It provides a natural source of type I collagen, which is the most abundant form of collagen found in the human body. The product is designed to meet the demands of various industries, including biomedical, cosmetic, and pharmaceutical applications.
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Lab products found in correlation
2 protocols using nutragen type 1 bovine collagen
1
Migration of Monocytes into Collagen Matrices
2
Migration of Monocytes into Collagen Matrices
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Collagen matrices were prepared from Nutragen Type I Bovine Collagen (Advanced BioMatrix) in 1X MEM Eagle supplemented with 7.5% sodium bicarbonate (pH 9) in optical-grade black 96-well plates. Matrices were allowed to polymerize at 37°C for 60 min to overnight, and then equilibrated ≥ 60 min in monocyte growth media supplemented with 3% serum. In each well, 5×104 monocytes or macrophages pre-labeled with Cell Trace CFSE (Invitrogen) and pre-infected with T. gondii (THP-1: MOI 4.5 with ME49 mCherry, 4 h; RAW 264.7: MOI 3 with ME49 mCherry) were allowed to migrate into the matrix. After overnight migration at 37°C and 5% CO2, the samples were rinsed with PBS and fixed with 4% formaldehyde. Z-stack image series of the monocytes captured within the collagen matrices were captured with the above-described spinning disk microscopy system and an EC Plan-Neofluar 10x (N.A. 0.30) objective. Volocity software was used to generate 3-D renderings of the Z-stacks and locate each monocyte using an automated Object Finding protocol based on fluorescence intensity and cell size. Vertical distance migrated was calculated as the distance between the centroid of the cell with the highest Z-position and every other cell.
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