High-precision three-dimensional inkjet technology for live cell bioprinting

Daisuke Takagi, Waka Lin, Takahiko Matsumoto, Hidekazu Yaginuma, Natsuko Hemmi, Shigeo Hatada, Manabu Seo

Article ID: 208
Vol 5, Issue 2, 2019, Article identifier:208

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In recent years, bioprinting has emerged as a promising technology for the construction of three-dimensional (3D) tissues to be used in regenerative medicine or in vitro screening applications. In the present study, we present the development of an inkjet-based bioprinting system to arrange multiple cells and materials precisely into structurally organized constructs. A novel inkjet printhead has been specially designed for live cell ejection. Droplet formation is powered by piezoelectric membrane vibrations coupled with mixing movements to prevent cell sedimentation at the nozzle. Stable drop-on-demand dispensing and cell viability were validated over an adequately long time to allow the fabrication of 3D tissues. Reliable control of cell number and spatial positioning was demonstrated using two separate suspensions with different cell types printed sequentially. Finally, a process for constructing stratified Mille-Feuille-like 3D structures is proposed by alternately superimposing cell suspensions and hydrogel layers with a controlled vertical resolution. The results show that inkjet technology is effective for both two-dimensional patterning and 3D multilayering and has the potential to facilitate the achievement of live cell bioprinting with an unprecedented level of precision.


Drop-on-demand; three-dimensional tissue engineering; drug discovery; regenerative medicine; hydrogel

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Takahashi K, Tanabe K, Ohnuki M, et al., 2007, Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell, 131(5):861-72. DOI 10.1016/j. cell.2007.11.019.

Groll J, Boland T, Blunk T, et al., 2016, Biofabrication: Reappraising the Definition in an Evolving Field. Biofabrication, 8(1):13001-6. DOI 10.1088/1758- 5090/8/1/013001.

Murphy SV, Atala A, 2014, 3D Bioprinting of Tissues and Organs. Nat Biotechnol, 32(8):773-85. DOI 10.1038/ nbt.2958.

Arslan-Yildiz A, El Assal R, Chen P, et al., 2016, Towards Artificial Tissue Models: Past, Present, and Future of 3D Bioprinting. Biofabrication, 8(1):14103. DOI 10.1088/1758- 5090/8/1/014103.

Gudupati H, Dey M, Ozbolat I, 2016, A Comprehensive Review on Droplet-based Bioprinting: Past, Present and Future. Biomaterials, 102:20-42. DOI 10.1016/j. biomaterials.2016.06.012.

Derby B, 2008, Bioprinting: Inkjet Printing Proteins and Hybrid Cell-containing Materials and Structures. J Mater Chem, 18(47):5717-21. DOI 10.1039/b807560c.

Xu T, Jin J, Gregory C, et al., 2005, Inkjet Printing of Viable Mammalian Cells. Biomaterials, 26(1):93-9. DOI 10.1016/j. biomaterials.2004.04.011.

Nakamura M, Kobayashi A, Takagi F, et al., 2005, Biocompatible Inkjet Printing Technique for Designed Seeding of Individual Living Cells. Tissue Eng, 11(11-12):1658-66. DOI 10.1089/ten.2005.11.1658.

Saunders RE, Gough JE, Derby B, 2008, Delivery of Human Fibroblast Cells by Piezoelectric Drop-on-demand Inkjet Printing. Biomaterials, 29(2):193-203. DOI 10.1016/j. biomaterials.2007.09.032.

Gross A, Schöndube J, Niekrawitz S, et al., 2013, Single-cell Printer: Automated, on Demand, and Label Free. J Lab Autom, 18(6):504-18. DOI 10.1177/2211068213497204.

Cheng E, Yu H, Ahmadi A, et al., 2016, Investigation of the Hydrodynamic Response of Cells in Drop on Demand Piezoelectric Inkjet Nozzles. Biofabrication, 8(1):15008. DOI 10.1088/1758-5090/8/1/015008.

Herran CL, Huang Y, Chai W, 2012, Performance Evaluation of Bipolar and Tripolar Excitations During Nozzle-jetting-based Alginate Microsphere Fabrication. J Micromech Microeng, 22(8):85025. DOI 10.1088/0960-1317/22/8/085025.

Kim YK, Park JA, Yoon WH, et al., 2016, Drop-on-demand Inkjet-based Cell Printing with 30-μm Nozzle Diameter for Cell-level Accuracy. Biomicrofluidics, 10(6):064110. DOI 10.1063/1.4968845.

Arai K, Iwanaga S, Toda H, et al., 2011, Three-dimensional Inkjet Biofabrication Based on Designed Images. Biofabrication, 3(3):34113. DOI 10.1088/1758-5082/3/3/034113.

Cui X, Boland T, 2009, Human Microvasculature Fabrication using Thermal Inkjet Printing Technology. Biomaterials, 30(31):6221-7. DOI 10.1016/j.biomaterials.2009.07.056.

Faulkner-Jones A, Fyfe C, Cornelissen DJ, et al., 2015, Bioprinting of Human Pluripotent Stem Cells and their Directed Differentiation into Hepatocyte-like Cells for the Generation of Mini-livers in 3D. Biofabrication, 7(4):44102. DOI 10.1088/1758-5090/7/4/044102.

Nakamura M, Iwanaga S, Henmi C, et al., 2010, Biomatrices and Biomaterials for Future Developments of Bioprinting and Biofabrication. Biofabrication, 2(1):14110. DOI 10.1088/1758-5082/2/1/014110.

Perçin G, Khuri-Yakub BT, 2003, Piezoelectric Droplet Ejector for Ink-jet Printing of Fluids and Solid Particles. Rev Sci Instrum, 74(2):1120-7. DOI 10.1063/1.1532839.

Zhang Z, Chai W, Xiong R, et al., 2017, Printing-induced Cell Injury Evaluation During Laser Printing of 3T3 Mouse Fibroblasts. Biofabrication, 9(2):25038. DOI 10.1088/1758-5090/aa6ed9.

Hu W, Berdugo C, Chalmers JJ, 2011, The Potential of Hydrodynamic Damage to Animal Cells of Industrial Relevance: Current Understanding. Cytotechnology, 63(5):445-60. DOI 10.1007/s10616-011-9368-3.

Moon S, Hasan SK, Song YS, et al., 2010, Layer by Layer Three-dimensional Tissue Epitaxy by Cell-laden Hydrogel Droplets. Tissue Eng Part C Methods, 16(1):157-66. DOI 10.1089/ten.tec.2009.0179.

Koch L, Deiwick A, Schlie S, et al., 2012, Skin Tissue Generation by Laser Cell Printing. Biotechnol Bioeng, 109(7):1855-63.

Malda J, Visser J, Melchels FP, et al., 2013, 25th Anniversary Article: Engineering Hydrogels for Biofabrication. Adv Mater, 25(36):5011-28. DOI 10.1002/adma.201302042.

Jungst T, Smolan W, Schacht K, et al., 2016, Strategies and Molecular Design Criteria for 3D Printable Hydrogels. Chem Rev, 116(3):1496-539. DOI 10.1021/acs. chemrev.5b00303.

DOI: http://dx.doi.org/10.18063/ijb.v5i2.208


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