Fig. 2.

Fig. 2. Apple-derived cellulose scaffold generation for 3D mammalian cell culture. McIntosh red apples were subjected to a controlled drying process at 220°C for up to 5 min to rigidify the outer hypanthium tissue. Following this, the apples underwent precise slicing using a mandolin slicer, ensuring thin and uniform sections while removing the cores. Subsequently, uniform segments measuring 2.0 × 0.5 cm were excised from the sliced apples and individually placed in microcentrifuge tubes for further processing. Decellularization of these apple segments ensued, involving the removal of cellular components while preserving the structural integrity of the cellulose matrix. Post-decellularization, the segments underwent surface modification with various chemistries. These modifications included coating with Type 1 collagen, chemical cross-linking using glutaraldehyde, or incubation in phosphate-buffered saline (PBS).The treated segments, now transformed into scaffolds, were introduced into mammalian cell culture medium (DMEM) and incubated for 12 h under standard tissue culture conditions (37°C, 5% CO₂). This incubation period allowed for equilibration and preparation of the scaffolds for subsequent cell seeding. For cell seeding, polydimethylsiloxane (PDMS)-coated 24-well plates were utilized, with each well containing 40 ml of cell suspension. The seeded scaffolds were incubated for 6 h to facilitate cell attachment and initial colonization. Subsequently, the wells were filled with DMEM and maintained for up to 12 weeks, providing an extended culture period to assess cell growth, viability, and functionality within the scaffolds. (Reproduced, with permission, copyright 2014, PLOS).

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