be superimposed to build a 3D construct of the desired
geometry.
5. Gently dispense few mL of the crosslinking solution on the
construct surface to induce alginate crosslinking.
6. After 5 min, remove the crosslinking solution, and gently wash
the crosslinked scaffold in cell culture medium to terminate the
gelation process.
7. Check the constructs under an inverted microscope. Two representative cell-laden constructs with different printing patterns are reported in Fig. 2.
8. Incubate constructs at 37
C in a 5% CO 2 atmosphere with
frequent medium changes (see Note 7). At selected timepoints, constructs can be retrieved and used for biological
characterization.
Fig. 1 Workflow for the fabrication of 3D constructs via direct-write deposition of thermosensitive hydrogel:
(1) 3D printing apparatus, (2) syringe loading and sol-gel transition, (3) scaffold manufacturing and
crosslinking
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Sara Maria Giannitelli et al.
geometry.
5. Gently dispense few mL of the crosslinking solution on the
construct surface to induce alginate crosslinking.
6. After 5 min, remove the crosslinking solution, and gently wash
the crosslinked scaffold in cell culture medium to terminate the
gelation process.
7. Check the constructs under an inverted microscope. Two representative cell-laden constructs with different printing patterns are reported in Fig. 2.
8. Incubate constructs at 37
C in a 5% CO 2 atmosphere with
frequent medium changes (see Note 7). At selected timepoints, constructs can be retrieved and used for biological
characterization.
Fig. 1 Workflow for the fabrication of 3D constructs via direct-write deposition of thermosensitive hydrogel:
(1) 3D printing apparatus, (2) syringe loading and sol-gel transition, (3) scaffold manufacturing and
crosslinking
140
Sara Maria Giannitelli et al.