Rat AEC Type II (ATCC, L2 cell line # CCL-149™) were used. L2 cells at passage 7 were seeded onto six-well BioFlex Collagen I coated plates (Flexcell International Corporation, Burlington, NC, United States) at a density of 1.5 × 105 cells/well in Dulbecco’s modified Eagle medium (DMEM, with High Glucose, L-glutamine, Phenol Red and Sodium Pyruvate, Gibco, # 11995073) supplemented with 10% fetal bovine serum (FBS, Gibco) and 50 μg/mL gentamycin sulfate (Gibco, # 15710049). Cells were incubated at 37°C, 21% O2 and 6.5% CO2 for 24 h. Sixteen hours before stretch experiments, cells were washed twice with sterile phosphate-buffered saline (PBS) and then incubated with DMEM (without Phenol Red, Merck Sigma-Aldrich, #D1145) containing 1% FBS, 50 µg gentamycin sulfate/mL, 4 mM L-alanyl-L-glutamine (Merck Sigma-Aldrich, # G8541), and 1mM Sodium Pyruvate (Gibco, # 11360070).
Bioflex collagen 1
Manufactured by Flexcell
Sourced in United States
The BioFlex Collagen I is a laboratory equipment product designed for cell culture applications. It provides a collagen-coated substrate for cell attachment and growth. The core function of this product is to offer a suitable surface for culturing cells in an in vitro environment.
Automatically generated - may contain errors
Lab products found in correlation
Gentamycin sulfate, by Merck Group (1 mentions)
Phenol red, by Thermo Fisher Scientific (1 mentions)
Gentamycin sulfate, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Phenol red, by Merck Group (1 mentions)
L alanyl l glutamine, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Fx 5000, by Flexcell (1 mentions)
2 protocols using bioflex collagen 1
1
Rat Alveolar Epithelial Cell Culture
2
Biaxial Strain Induces FAK Activation
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The biaxial strain was performed essentially as described elsewhere (Uzer et al., 2015 (link)). In brief, REF52 intact cells and cytoplasts were plated at a density of 30,000 cell/cm2 per well in six-well BioFlex collagen-I coated plates (BF-3001C; Flexcell). After plating for either 3 or 18 h, cells were subjected to dynamic uniform biaxial cyclic strain at 5% magnitude at 10 cpm for 20 min using the Flexcell FX 5000 under conditions of 37°C and 5% CO2. Control plates were handled the same but without strain application. Immediately after strain, whole-cell lysates were prepared and probed for phospho-FAK, total FAK, and GAPDH via Western blot analysis. Blots were analyzed using ImageJ. Data were derived from three independent experiments.
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