| Bioactivity |
1. Cultivation of BHK-21 and 2BS cell line BHK-21 or 2BS cells were plated onto the bottom of the 96-well cell culture plates at the density of 1x10e3cells/well, respectively, with 0.1ml RPMI1640 contained 10% fetal bovine serum in the well. BHK-21 or 2BS cells were incubated in standard conditions (at 37 degree C with 5% CO2 in a humidified incubator) for 24 hours. 2. Biochemical activity analysis of Human-like Collagen) Complete culture media was removed after 24 hours, rHuCollagen was added at various concentrations: 0.01%, 0.03%, 0.05%, 0.07%, 0.09%, 0.10%, 0.11%, 0.13% and a complete medium was used as blank control. Cells were incubated in standard conditions (at 37 degree C with 5% CO2 in a humidified incubator). The morphology of cell was observed by reverse microscope after 24hr, 48hr, 72hr, 96hr, and 120hr. Thereafter, the viable cell count was determined by a colorimetric method using MTT (3-4,5-dimethylthiazol-2yl-2, 5-diphenyl tetrazolium bromide) after 24hr, 48hr, 72hr, 9
|