3. Example: growth of HepG2 hepatocyte cell line in Matrigel™ coated Alvetex Scaffold
3.1. Cell Culture details
HepG2 cells (ATCC, HB-8065) were routinely maintained in T-75 flasks. HepG2 complete media consisted of: MEM media (Gibco (Thermo Fisher), 21090) supplemented with 10 % v/v FBS, 2 mM L-glutamine and 100 U/mL Penicillin/ Streptomycin. Alvetex Scaffold 6-well inserts (AVP004) in 6-well plates, were coated with Matrigel™ as described above.
Cells were seeded at a density of 1 × 106 cells in 150 μL media suspension per disc and were left to settle for 60 minutes in an incubator (5 % CO2, 37 °C). Media was carefully added to each sample well (9 mL per well). Cultures were maintained for 7 days, with media changed on days 3 and 5.
3.2. Results
Pre-coating of Alvetex Scaffold discs with Matrigel™ resulted in enhanced infiltration of cells into the scaffold compared with control cultures in untreated Alvetex Scaffold. Cells were seen to occupy the entire depth of the scaffold after 7 days of growth in Matrigel™ coated discs, while cells grown in untreated Alvetex Scaffold occupied only the upper half of the scaffold. These findings indicate that pre-treatment of Alvetex Scaffold with extracellular matrix products is able to enhance the growth of appropriate cell types into the 3D structure.
