2.6 Cell culture study
The HUVECs were cultured as per the manufacturer’s procedure in RPMI
1640 cell culture medium, which consisted of 10% fetal bovine serum,
0.292 mg/mL L-glutamine, 4.766 mg/mL HEPES, 0.85 mg/mL
NaHCO3, 1% penicillin (100 units/mL), and streptomycin
(100 μg/mL). The cells were maintained at 37 °C in a humid atmosphere
containing 5% CO2. The culture medium was replaced
every day, after cultured for the first three days, and the cells then
suspended in fresh medium. The final suspension density of 6 ×
106 cells/mL was prepared. The samples of SA/Gel and
A-SA-Gel hydrogel scaffolds were sterilized by 75% ethanol for 6 h
under UV light and then rinsed three times with sterilized PBS. Next,
the scaffolds were immersed in a fresh cell culture medium for 2 h
before cell seeding. Finally, the cell suspension was lightly inoculated
on the treated A-SA-Gel hydrogel scaffold until the surface of the
scaffold was fully covered. After 4 h of initial cell attachment, the
culture medium was added gently until the whole scaffolds were
completely immersed. The experiments were conducted under static cell
culture conditions and the cell culture medium was changed every day.
Moreover, the prepared SA/Gel scaffold without albumen as a control
group.