Example 7
Fibroblast cells were expanded in a growth medium containing 90% DMEM, 10% fetal bovine serum (FBS), 100 U/ml penicillin, 1000 U/ml streptomycin. Cell cultures were maintained at 37° C. in an incubator with 95% air and 5% CO2. The cultures were replenished with fresh medium at 37° C. every two days. For adhesion, cells were seeded on silk films that were pre-cast in 24-well plates with 50,000 cells per well in 1 ml of serum-containing medium. Empty wells with tissue culture plastic (TCP) and no silk served as controls. Cell attachment was evaluated 3 hr after cell seeding by adding 50 μl of alamar blue to the culture medium, culturing for another 6 hr, and determining the medium fluorescence (Ex=560 nm, Em=590 nm). During the culture, cell proliferation was determined using alamar blue staining and cell morphology was monitored by phase contrast light microscopy (Carl Zeiss, Inc., Jena, Germany).
All experiments were performed with a minimum of N=3 for each data point. Statistical analysis was performed by one-way analysis of variance (ANOVA) and Student-Newman-Keuls Multiple Comparisons Test. Differences were considered significant when p≤0.05, and very significant when p≤0.01.