High-precision 3D inkjet technology for live cell bioprinting

VIEWS - 212 (Abstract) 125 (PDF)
Daisuke Takagi, Waka Lin, Takahiko Matsumoto, Hidekazu Yaginuma, Natsuko Hemmi, Shigeo Hatada, Manabu Seo


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; 3D tissue engineering, drug discovery, regenerative medicine, and hydrogel

Full Text:



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.


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-


Murphy SV, Atala A, 2014, 3D Bioprinting of Tissues

and Organs. Nat Biotechnol, 32(8):773-85. DOI 10.1038/


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-


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.


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.


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.


Gross A, Schöndube J, Niekrawitz S, et al., 2013, Singlecell

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,

(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):34113. DOI 10.1088/1758-5082/3/3/034113.

Cui X, Boland T, 2009, Human Microvasculature Fabrication

using Thermal Inkjet Printing Technology. Biomaterials,

(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


Hu W, Berdugo C, Chalmers JJ, 2011, The Potential of

Hydrodynamic Damage to Animal Cells of Industrial

Relevance: Current Understanding. Cytotechnology,

(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.


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


  • There are currently no refbacks.

Copyright (c) 2019 Daisuke Takagi, Waka Lin, Takahiko Matsumoto, Hidekazu Yaginuma, Natsuko Hemmi, Shigeo Hatada, Manabu Seo

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.