Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

Figure 3

Cell proliferation and viability of nasal chondrocytes isolated using digest protocols of 1 h or 12 h with 750 or 3000 U/ml of collagenase enzyme with physical agitation and subsequent culture in alginate beads for 21 days. (a) DNA content normalized to wet weight (ng/mg) at day 0 and day 21. (b) Live/dead cell viability at day 0 and day 21. (c) Cell density (per cm²) for peripheral and core regions. (d) Hematoxylin and eosin (H&E) staining of cell morphology in alginate bead constructs at day 0 imaged under high magnification. Scale bar: 20 µm (1 h) and 10 µm (12 h) and gross alginate bead constructs at day 21. Scale bar: 1 mm. (e) Percentage chondron per construct (%) determined using ImageJ analysis. to 1 h incubation for the same enzyme concentration and to 750 U/ml enzyme concentration for the same incubation period .

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