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Briefly, fibrillar collagen preparations were made from Vitrogen 100 bovine type I collagen (Cohesion Technologies, Inc., Palo Alto, CA, USA) in accordance with the manufacturer's instructions.
Briefly, purified type I collagen was purchased as a solution of acid-stabilized monomer (Vitrogen; Cohesion Technologies, Inc., Palo Alto, CA).
The supernatant was then centrifuged, and the alveolar type II (ATII) cell-enriched pellet was resuspended in complete medium and seeded onto type I collagen-coated plastic (Vitrogen 100; Cohesion Technologies, Palo Alto, CA).
The cell suspensions were mixed with 100 μl of a buffered collagen solution (pH 7.4), containing 1.67 mg/ml bovine collagen type I (Invitrogen, Cohesion Technologies, Palo Alto, CA).
In brief, a suspension of 2.5 × 105 leukocytes or 8 × 104 SH-SY5Y cells or PC-3 cells in 50 μl RPMI was mixed with 100 μl of a buffered collagen solution (pH 7.4), containing 1.67 mg/ml bovine collagen type I (Invitrogen, Cohesion Technologies, Palo Alto, CA).
Recombinant human vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and anti-VEGF (MAB293) were purchased from R&D Systems Inc. (Minneapolis, MN); adult human serum-AB from Gemini Bioproducts (Woodland, CA); bovine skin-derived type I collagen from Cohesion Technologies, Inc. (Palo Alto, CA).
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