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Kenichi Arai, Yoshinari Tsukamoto, Hirotoshi Yoshida, et al.

Figure 8. The culture of fabricated 3D Alg-Ph/Gelatin-Ph gel sheets including living cells by 3D-bioprinter at day 1. The fabricated
3D Alg-Ph/Gelatin-Ph gel sheets (A), the photomicrograph (B), and phase contrast image (C) of fibroblasts in Alg-Ph/Gelatin-Ph gel.
Fibroblasts were stained with Hoechst 33342 and Alexa Fluor 488 phalloidin. Fibroblasts were observed at high magnification
(D–F).

Figure 9. The culture of fabricated 3D Alg-Ph/Gelatin-Ph gel sheets including living cells by 3D biopriter at day 7. The fabricated
3D Alg-Ph/Gelatin-Ph gel sheet (A), the photomicrograph (B), and phase contrast image (C) of fibroblasts in Alg-Ph/Gelatin-Ph gel.
Fibroblasts were stained with Hoechst 33342 and Alexa Fluor 488 phalloidin. Fibroblasts were observed at high magnification
(D–F).

structures in vivo. It is well known that tissues and ginate hydrogel has been used mainly because of its
organs have specific properties and components such good fabricating property. However, this fabrication
as capillary vessels, micro-structures, multi-cells, cell technique is still problematic and show several ob-
interactions, and extracellular matrices (ECMs). How- stacles that have to be resolved. Firstly, the fabricated
ever, it is difficult to reconstruct artificial tissues and alginate gel structures cannot maintain its morphology
organs with these tissue specific properties in vitro. for a long time. Crosslinking through calcium ion in
We recently reported the fabrication of 3D alginate gel the alginate hydrogel show gradual weakness due to a
structures with micro-structures and multi-cell proper- low concentration of calcium ion in the medium. Se-
ties by using the 3D-bioprinter [14–15] . 3D alginate gel condly, alginate hydrogel is not suitable for tissue re-
structures had been fabricated by crosslinking the cal- generation due to its poor cell-adhesive property. In
cium ion and carboxyl group of sodium alginate. Al- general, cell adhesion and extension is crucial for the

International Journal of Bioprinting (2016)–Volume 2, Issue 2 159