Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids

Kai Ling; Guoyou Huang; Juncong Liu; Xiaohui Zhang; Yufei Ma; Tianjian Lu; Feng Xu

doi:10.15302/J-ENG-2015062

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Engineering ›› 2015, Vol. 1 ›› Issue (2) : 269-274. DOI: 10.15302/J-ENG-2015062

Research

Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids

Kai Ling¹^,² ,
Guoyou Huang²^,³ ,
Juncong Liu² ,
Xiaohui Zhang²^,³ ,
Yufei Ma²^,³ ,
Tianjian Lu² ,
Feng Xu²^,³

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Abstract

Cellular spheroids serving as three-dimensional (3D) in vitro tissue models have attracted increasing interest for pathological study and drug-screening applications. Various methods, including microwells in particular, have been developed for engineering cellular spheroids. However, these methods usually suffer from either destructive molding operations or cell loss and non-uniform cell distribution among the wells due to two-step molding and cell seeding. We have developed a facile method that utilizes cell-embedded hydrogel arrays as templates for concave well fabrication and in situ MCF-7 cellular spheroid formation on a chip. A custom-built bioprinting system was applied for the fabrication of sacrificial gelatin arrays and sequentially concave wells in a high-throughput, flexible, and controlled manner. The ability to achieve in situ cell seeding for cellular spheroid construction was demonstrated with the advantage of uniform cell seeding and the potential for programmed fabrication of tissue models on chips. The developed method holds great potential for applications in tissue engineering, regenerative medicine, and drug screening.

Keywords

MCF-7 cellular spheroids / bioprinting / hydrogels / concave wells / tissue on a chip

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Kai Ling, Guoyou Huang, Juncong Liu, Xiaohui Zhang, Yufei Ma, Tianjian Lu, Feng Xu. Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids. Engineering, 2015, 1(2): 269‒274 https://doi.org/10.15302/J-ENG-2015062

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	T. M. Achilli, J. Meyer, J. R. Morgan. Advances in the formation, use and understanding of multi-cellular spheroids. Expert Opin. Biol. Ther., 2012, 12(10): 1347−1360

[2]	M. Rimann, U. Graf-Hausner. Synthetic 3D multicellular systems for drug development. Curr. Opin. Biotechnol., 2012, 23(5): 803−809

[3]	L. Wang, Engineering three-dimensional cardiac microtissues for potential drug screening applications. Curr. Med. Chem., 2014, 21(22): 2497−2509

[4]	J. Rouwkema, J. de Boer, C. A. van Blitterswijk. Endothelial cells assemble into a 3-dimensional prevascular network in a bone tissue engineering construct. Tissue Eng., 2006, 12(9): 2685−2693

[5]	E. Fennema, N. Rivron, J. Rouwkema, C. van Blitterswijk, J. de Boer. Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol., 2013, 31(2): 108−115

[6]	K. Takayama, 3D spheroid culture of hESC/hiPSC-derived hepatocyte-like cells for drug toxicity testing. Biomaterials, 2013, 34(7): 1781−1789

[7]	P. R. Baraniak, T. C. McDevitt. Scaffold-free culture of mesenchymal stem cell spheroids in suspension preserves multilineage potential. Cell Tissue Res., 2012, 347(3): 701−711

[8]	A. P. Napolitano, Scaffold-free three-dimensional cell culture utilizing micromolded nonadhesive hydrogels. Biotechniques, 2007, 43(4): 494, 496−500

[9]	D. M. Dean, J. R. Morgan. Cytoskeletal-mediated tension modulates the directed self-assembly of microtissues. Tissue Eng. Part A, 2008, 14(12): 1989−1997

[10]	J. Liu, Soft fibrin gels promote selection and growth of tumorigenic cells. Nat. Mater., 2012, 11(8): 734−741

[11]	J. Friedrich, C. Seidel, R. Ebner, L. A. Kunz-Schughart. Spheroid-based drug screen: Considerations and practical approach. Nat. Protoc., 2009, 4(3): 309−324

[12]	H. F. Chan, Y. Zhang, Y. P. Ho, Y. L. Chiu, Y. Jung, K. W. Leong. Rapid formation of multicellular spheroids in double-emulsion droplets with controllable microenvironment. Sci. Rep., 2013, 3: 3462

[13]	F. Langenbach, Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering. Nat. Protoc., 2011, 6(11): 1726−1735

[14]	M. Inamori, H. Mizumoto, T. Kajiwara. An approach for formation of vascularized liver tissue by endothelial cell-covered hepatocyte spheroid integration. Tissue Eng. Part A, 2009, 15(8): 2029−2037

[15]	S. F. Wong, D. Y. No, Y. Y. Choi, D. S. Kim, B. G. Chung, S. H. Lee. Concave microwell based size-controllable hepatosphere as a three-dimensional liver tissue model. Biomaterials, 2011, 32(32): 8087−8096

[16]	D. Huh, B. D. Matthews, A. Mammoto, M. Montoya-Zavala, H. Y. Hsin, D. E. Ingber. Reconstituting organ-level lung functions on a chip. Science, 2010, 328(5986): 1662−1668

[17]	D. Huh, Y. S. Torisawa, G. A. Hamilton, H. J. Kim, D. E. Ingber. Microengineered physiological biomimicry: Organs-on-chips. Lab Chip, 2012, 12(12): 2156−2164

[18]	G. Wang, Modeling the mitochondrial cardiomyopathy of Barth syndrome with induced pluripotent stem cell and heart-on-chip technologies. Nat. Med., 2014, 20(6): 616−623

[19]	R. A. Rezende, Scalable biofabrication of tissue spheroids for organ printing. Procedia CIRP, 2013, 5: 276−281

[20]	R. J. Thomas, The effect of three-dimensional co-culture of hepatocytes and hepatic stellate cells on key hepatocyte functions in vitro. Cells Tissues Organs (Print), 2005, 181(2): 67−79

[21]	Y. C. Tung, A. Y. Hsiao, S. G. Allen, Y. S. Torisawa, M. Ho, S. Takayama. High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. Analyst, 2011, 136(3): 473−478

[22]	G. R. Souza, Three-dimensional tissue culture based on magnetic cell levitation. Nat. Nanotechnol., 2010, 5(4): 291−296

[23]	T. Liu, M. Winter, B. Thierry. Quasi-spherical microwells on superhydrophobic substrates for long term culture of multicellular spheroids and high throughput assays. Biomaterials, 2014, 35(23): 6060−6068

[24]	S. E. Yeon, Application of concave microwells to pancreatic tumor spheroids enabling anticancer drug evaluation in a clinically relevant drug resistance model. PLoS ONE, 2013, 8(9): e73345

[25]	L. Kang, M. J. Hancock, M. D. Brigham, A. Khademhosseini. Cell confinement in patterned nanoliter droplets in a microwell array by wiping. J. Biomed. Mater. Res. A, 2010, 93(2): 547−557

[26]	H. Tekin, M. Anaya, M. D. Brigham, C. Nauman, R. Langer, A. Khademhosseini. Stimuli-responsive microwells for formation and retrieval of cell aggregates. Lab Chip, 2010, 10(18): 2411−2418

[27]	C. Kim, J. H. Bang, Y. E. Kim, S. H. Lee, J. Y. Kang. On-chip anticancer drug test of regular tumor spheroids formed in microwells by a distributive microchannel network. Lab Chip, 2012, 12(20): 4135−4142

[28]	C. Kim, 3-Dimensional cell culture for on-chip differentiation of stem cells in embryoid body. Lab Chip, 2011, 11(5): 874−882

[29]	H. C. Moeller, M. K. Mian, S. Shrivastava, B. G. Chung, A. Khademhosseini. A microwell array system for stem cell culture. Biomaterials, 2008, 29(6): 752−763

[30]	Y. S. Hwang, B. G. Chung, D. Ortmann, N. Hattori, H. C. Moeller, A. Khademhosseini. Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11. Proc. Natl. Acad. Sci. U.S.A., 2009, 106(40): 16978−16983

[31]	Y. Xia, G. M. Whitesides. Soft lithography. Annu. Rev. Mater. Sci., 1998, 28(1): 153−184

[32]	Y. Y. Choi, B. G. Chung, D. H. Lee, A. Khademhosseini, J. H. Kim, S. H. Lee. Controlled-size embryoid body formation in concave microwell arrays. Biomaterials, 2010, 31(15): 4296−4303

[33]	A. Y. Hsiao, Microfluidic system for formation of PC-3 prostate cancer co-culture spheroids. Biomaterials, 2009, 30(16): 3020−3027

Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (11372243, 11532009, and 11522219), the China Postdoctoral Science Foundation (2013M540742), the Doctoral Program of Higher Education of China (20130201120071), the Natural Science Basic Research Plan in Shaanxi Province of China (2014JQ1004), and the Fundamental Research Funds for the Central Universities.

Compliance with ethics guidelines

Kai Ling, Guoyou Huang, Juncong Liu, Xiaohui Zhang, Yufei Ma, Tianjian Lu, and Feng Xu declare that they have no conflict of interest or financial conflicts to disclose.

Funding