Formation of Cell Aggregates Using Microfabricated Hydrogel Chambers for Assembly into Larger Tissues
Masaki Iwase, Masumi Yamada, Emi Yamada,
and Minoru Seki
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan
This paper presents a fabrication process for cell aggregates with controlled shapes that can be used as building units for constructing relatively large tissue models. Microfabricated hydrogel-based chambers with non-adhesive surface characteristics were prepared via a micromolding process. Alginate was used as the hydrogel matrix, which facilitated the efficient formation of aggregates from cells retained inside the microchamber. We employed several types of toroidal and lattice-shaped hydrogel microchambers with different geometries. We examined the effect of cell type on the aggregate formation process using NIH-3T3, C2C12, and HepG2 cells and clearly observed that aggregation behavior is highly dependent on cell type. In addition, we tried to construct 2-layered capillarylike tissues by stacking heterotypic toroidal cell aggregates, which mimic blood vessels. The presented cell aggregate-based tissue fabrication process could become a versatile approach for preparing complex and scaffold-free 3D tissue models.
and Minoru Seki, “Formation of Cell Aggregates Using Microfabricated Hydrogel Chambers for Assembly into Larger Tissues,” J. Robot. Mechatron., Vol.25, No.4, pp. 682-689, 2013.
-  Y. Zheng, J. Chen, M. Craven, N. W. Choi, S. Totorica, A. Diaz-Santana, P. Kermani, B. Hempstead, C. Fischbach-Teschl, J. A. López, and A. D. Stroock, “In vitro Microvessels for the Study of Angiogenesis and Thrombosis,” Proc. of the National Academy of Sciences of the United States of America, Vol.109, No.24, pp. 9342-9347, 2012.
-  A. Ivascu and M. Kubbies, “Rapid Generation of Single-Tumor Spheroids for High-throughput Cell Function and Toxicity Analysis,” J. of Biomolecular Screening, Vol.11, No.8, pp. 922-932, 2006.
-  J. Yang, M. Yamato, T. Shimizu, H. Sekine, K. Ohashi, M. Kanzaki, T. Ohki, K. Nishida, and T. Okano, “Reconstruction of Functional Tissues with Cell Sheet Engineering,” Biomaterials, Vol.28, No.34, pp. 5033-5043, 2007.
-  J. He, Y. Liu, X. Hao, M. Mao, L. Zhu, and D. Li, “Bottom-up Generation of 3D Silk Fibroin-Gelatin Microfluidic Scaffolds with Improved Structural and Biological Properties,” Materials Letters, Vol.78, pp. 102-105, 2012.
-  M. I. Santos, K. Tuzlakoglu, S. Fuchs, M. E. Gomes, K. Peters, R. E. Unger, E. Piskin, R. L. Reis, and C. J. Kirkpatrick, “Endothelial Cell Colonization and Angiogenic Potential of Combined Nanoand Micro-fibrous Scaffolds for Bone Tissue Engineering,” Biomaterials, Vol.29, No.32, pp. 4306-4313, 2008.
-  S. Moon, S. K. Hasan, Y. S. Song, F. Xu, H. O. Keles, F. Manzur, S. Mikkilineni, J. W. Hong, J. Nagatomi, E. Haeggstrom, A. Khademhosseini, and U. Demirci, “Layer by Layer Three-Dimensional Tissue Epitaxy by Cell-Laden Hydrogel Droplets,” Tissue Engineering-Part C: Methods, Vol.16, No.1, pp. 157-166, 2010.
-  Y. T. Matsunaga, Y. Morimoto, and S. Takeuchi, “Molding Cell Beads for Rapid Construction of Macroscopic 3D Tissue Architecture,” Advanced Materials, Vol.23, No.12, pp. H90-94, 2011.
-  M. Yamada, S. Sugaya, Y. Naganuma, and M. Seki, “Microfluidic Synthesis of Chemically and Physically Anisotropic Hydrogel Microfibers for Guided Cell Growth and Networking,” Soft Matter, Vol.8, No.11, pp. 3122-3130, 2012.
-  M. Yamada, R. Utoh, K. Ohashi, K. Tatsumi, M. Yamato, T. Okano, and M. Seki, “Controlled Formation of Heterotypic Hepatic Microorganoids in Anisotropic Hydrogel Microfibers for Long-term Preservation of Liver-specific Functions,” Biomaterials, Vol.33, No.33, pp. 8304-8315, 2012.
-  M. Matsusaki, H. Ajiro, T. Kida, T. Serizawa, and M. Akashi, “Layer-by-layer Assembly through Weak Interactions and Their Biomedical Applications,” Advanced Materials, Vol.24, No.4, pp. 454-474, 2012.
-  C. Norotte, F. S. Marga, L. E. Niklason, and G. Forgacs, “Scaffoldfree Vascular Tissue Engineering Using Bioprinting,” Biomaterials, Vol.30, No.30, pp. 5910-5917, 2009.
-  T. Anada, T. Masuda, Y. Honda, J. Fukuda, F. Arai, T. Fukuda, and O. Suzuki, “Three-dimensional Cell Culture Device Utilizing Thin Membrane Deformation by Decompression,” Sensors and Actuators B: Chemical, Vol.147, No.1, pp. 376-379, 2010.
-  Y. Sakai and K. Nakazawa, “Technique for the Control of Spheroid Diameter Using Microfabricated Chips,” Acta Biomaterialia, Vol.3, No.6, pp. 1033-1040, 2007.
-  H. Ota and N. Miki, “Microfluidic Experimental Platform for Producing Size-controlled Three-dimensional Spheroids,” Sensors and Actuators A: Physical, Vol.169, No.2, pp. 266-273, 2011.
-  M. Okochi, T. Matsumura, and H. Honda, “Magnetic force-based Cell Patterning for Evaluation of the Effect of Stromal Fibroblasts on Invasive Capacity in 3D Cultures,” Biosensors and Bioelectronics, Vol.42, No.1, pp. 300-307, 2013.
-  J. Fukuda, A. Khademhosseini, Y. Yeo, X. Yang, J. Yeh, G. Eng, J. Blumling, C.-F. Wang, D. S. Kohane, and R. Langer, “Micromolding of Photocrosslinkable Chitosan Hydrogel for Spheroid Microarray and Co-cultures,” Biomaterials, Vol.27, No.30, pp. 5259-5267, 2006.
-  L. R. Garzoni, M. I. D. Rossi, A. P. D. N. de Barros, V. Guarani, M. Keramidas, L. B. L. Balottin, D. Adesse, C. M. Takiya, P. P. Manso, I. B. Otazu, M. d. N. Meirelles, and R. Borojevic, “Dissecting Coronary Angiogenesis: 3D Co-culture of Cardiomyocytes with Endothelial or Mesenchymal Cells,” Experimental Cell Research, Vol.315, No.19, pp. 3406-3418, 2009.
-  M. Kato-Negishi, Y. Tsuda, H. Onoe, and S. Takeuchi, “A Neurospheroid Network-Stamping Method for Neural Transplantation to the Brain,” Biomaterials, Vol.31, No.34, pp. 8939-8945, 2010.
-  K. Takayama, K. Kawabata, Y. Nagamoto, K. Kishimoto, K. Tashiro, F. Sakurai, M. Tachibana, K. Kanda, T. Hayakawa, M. K. Furue, and H. Mizuguchi, “3D Spheroid Culture of hESC/hiPSCderived Hepatocyte-like Cells for Drug Toxicity Testing,” Biomaterials, Vol.34, No.7, pp. 1781-1789, 2013.
-  S. Sakai, H. Inagaki, K. Inamoto, and M. Taya, “Wrapping Tissues with a Pre-established Cage-like Layer Composed of Living Cells,” Biomaterials, Vol.33, No.28, pp. 6721-6727, 2012.
-  N. Kojima, S. Takeuchi, and Y. Sakai, “Establishment of Selforganization System in Rapidly Formed Multicellular Heterospheroids,” Biomaterials, Vol.32, No.26, pp. 6059-6067, 2011.
-  C. M. Livoti and J. R. Morgan, “Self-Assembly and Tissue Fusion of Toroid-Shaped Minimal Building Units,” Tissue Engineering-Part A, Vol.16, No.6, pp. 2051-2061, 2010.
-  T. Masuda, N. Takei, T. Nakano, T. Anada, O. Suzuki, and F. Arai, “A Microfabricated Platform to Form Three-dimensional Toroidal Multicellular Aggregate,” Biomedical Microdevices, Vol.14, No.6, pp. 1085-1093, 2012.
-  K. Yoshimoto, R. Kojima, E. Takahashi, M. Ichino, H. Miyoshi, and Y. Nagasaki, “3D Cell Co-culture System on Hydrogel Micro-Patterned Surface Fabricated by Photolithography,” J. of Photopolymer Science and Technology, Vol.25, No.1, pp. 47-52, 2012.
-  T. A. Gwyther, J. Z. Hu, A. G. Christakis, J. K. Skorinko, S. M. Shaw, K. L. Billiar, and M. W. Rolle, “Engineered Vascular Tissue Fabricated from Aggregated Smooth Muscle Cells,” Cells Tissues Organs, Vol.194, No.1, pp. 13-24, 2011.
-  S. Sugaya, S. Kakegawa, S. Fukushima, M. Yamada, and M. Seki, “Micropatterning of Hydrogels on Locally Hydrophilized Regions on PDMS by Stepwise Solution Dipping and in Situ Gelation,” Langmuir, Vol.28, No.39, pp. 14073-14080, 2012.
-  N. W. Choi, M. Cabodi, B. Held, J. P. Gleghorn, L. J. Bonassar, and A. D. Stroock, “Microfluidic Scaffolds for Tissue Engineering,” Nature Materials, Vol.6, No.11, pp. 908-915, 2007.
-  J. Youssef, A. K. Nurse, L. B. Freund, and J. R. Morgan, “Quantification of the Forces Driving Self-Assembly of Three-dimensional Microtissues,” Proc. of the National Academy of Sciences of the United States of America, Vol.108, No.17, pp. 6993-6998, 2011.
-  K. Ohara, D. Kawakami, T. Takubo, Y. Mae, T. Tanikawa, A. Honda, and T. Arai, “Dextrous Cell Diagnosis Using Two-fingered Microhand with Micro Force Sensor,” J. of Micro-Nano Mechatronics, Vol.7, No.1-3, pp. 13-20, 2012.
-  Y. Yamanishi, T. Nakano, Y. Sawada, K. Itoga, T. Okano, and F. Arai, “Maskless Gray Scale Lithography and its 3D Microfluidic Applications,” J. of Robotics and Mechatronics, Vol.23, No.3, pp. 426-433, 2011.
-  M. Hagiwara, M. Niimi, T. Kawahara, Y. Yamanishi, H. Nakanishi, and F. Arai, “On-Chip Particle Sorting into Multiple Channels by Magnetically Driven Microtools,” J. of Robotics and Mechatronics, Vol.23, No.3, pp. 370-377, 2011.
-  A. Ichikawa, T. Tanikawa, S. Akagi, and K. Ohba, “Automatic Cell Cutting by High-Precision Microfluidic Control,” J. of Robotics and Mechatronics, Vol.23, No.1, pp. X13-18, 2011.
This article is published under a Creative Commons Attribution-NoDerivatives 4.0 International License.
Copyright© 2013 by Fuji Technology Press Ltd. and Japan Society of Mechanical Engineers. All right reserved.