2.6. Cell culture
Donor DPSCs (passage 6-7) were cultured in Alpha modification of Eagle’s
medium (α-MEM) supplemented with 10% FBS, 1% penicillin (100
U/mL)/streptomycin (100 µg/mL), 100 μM L-ascorbic acid 2-phosphate, and
2 mM L-glutamine at 37 °C in 5% CO2 humidified
atmosphere. Cells reaching 80–90% confluency were harvested using
0.05% (w/v) trypsin/ethylenediaminetetraacetic acid (EDTA) solution and
seeded onto scaffolds in 24-well culture plates (100 µL/well) at 1, 2,
4, 8, or 16×104 cells/sample. The scaffolds were
incubated for 3 hours at 37 °C with 5% CO2 to allow
diffusion of cells through the network of pores before adding culture
media (550 µL/well). DPSCs cultured on the surface of the well plate
(i.e. No scaffolds) were used as 2D control.