Design and Functional Testing of a Multichamber Perfusion Platform for Three-Dimensional Scaffolds
Figure 3
Cell culture experiments performed for the evaluation of the dynamic perfusion seeding. (1) Cell expansion: cell line MC3T3-E1 was cultured in standard T-flask in cell culture incubators (5% CO2, 37°C) and passaged when 60–70% confluence was reached (a). (2) Cell seeding: the seeding density was about 2 × 105 cells/50 μL of culture medium, and the scaffolds were porous PCL matrices with a porosity of 84%. Suspended cells were injected (b) or dropwise seeded (c, d) onto the scaffolds. Injection- (b) and dropwise-seeded (c) scaffolds were fixed within holder cartridges and incubated in a six-well plate for 6 hours. In addition, cell suspensions were poured on the top surface of each scaffold fixed in a holder cartridge within a perfusion culture chamber (d); then the bioreactor was closed (f), and incubated for 6 hours. (3) Analysis: formalin-fixed scaffolds were cut into 200 μm thick-longitudinal sections (g), and the distribution of the seeded cells within the thickness of the scaffold was observed via fluorescence microscopy after cell nuclei staining (h).
