AccScience Publishing / IJB / Volume 9 / Issue 1 / DOI: 10.18063/ijb.v9i1.630
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A 3D bioprinted tumor model fabricated with gelatin/sodium alginate/decellularized extracellular matrix bioink 

Jie Xu1† Shuangjia Yang1† Ya Su1 Xueyan Hu1 Yue Xi1 Yuen Yee Cheng2 Yue Kang3* Yi Nie4,5* Bo Pan6* Kedong Song1*
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1 State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China
2 Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology Sydney, NSW 2007, Australia
3 Department of Breast Surgery, Cancer Hospital of China Medical University, 44 Xiaoheyan Road, Dadong District, Shenyang 110042, China
4 Zhengzhou Institute of Emerging Industrial Technology, Zhengzhou 450000, China
5 Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
6 Department of Breast Surgery, The Second Hospital of Dalian Medical University, 467 Zhongshan Road, Shahekou District, Dalian, Liaoning 116023, China
Submitted: 29 May 2022 | Accepted: 2 September 2022 | Published: 28 October 2022
© 2022 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( )

Tissue-engineered scaffolds are more commonly used to construct three-dimension­al (3D) tumor models for in vitro studies when compared to the conventional two-dimensional (2D) cell culture because the microenvironments provided by the 3D tumor models closely resemble the in vivo system and could achieve higher success rate when the scaffolds are translated for use in pre-clinical animal model. Physical properties, heterogeneity, and cell behaviors of the model could be regulated to simulate different tumors by changing the components and concentrations of materials. In this study, a novel 3D breast tumor model was fabricated by bioprinting using a bioink that consists of porcine liver-derived decellularized extracellular matrix (dECM) with different concentrations of gelatin and sodium alginate. Primary cells were removed while extracellular matrix components of porcine liver were preserved. The rheolog­ical properties of biomimetic bioinks and the physical properties of hybrid scaffolds were investigated, and we found that the addition of gelatin increased hydrophilia and viscoelasticity, while the addition of alginate increased mechanical properties and porosity. The swelling ratio, compression modulus, and porosity could reach 835.43 ± 130.61%, 9.64 ± 0.41 kPa, and 76.62 ± 4.43%, respectively. L929 cells and the mouse breast tumor cells 4T1 were subsequently inoculated to evaluate biocompatibility of the scaffolds and to form the 3D models. The results showed that all scaffolds exhibited good biocompatibility, and the average diameter of tumor spheres could reach 148.52 ± 8.02 μm on 7 d. These findings suggest that the 3D breast tumor model could serve as an effective platform for anticancer drug screening and cancer research in vitro.

Tumor model
Decellularized extracellular matrix
Sodium alginate
Three-dimensional bioprinting

Ferlay J, Soerjomataram I, Dikshit R, et al., 2015, Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer, 136(5): E359–86.

Ferlay J, Colombet M, Soerjomataram I, et al., 2021, Cancer statistics for the year 2020: An overview. Int J Cancer, 149(8): 778–789.

Xin X, Yang H, Zhang F, et al., 2019, 3D cell coculture tumor model: A promising approach for future cancer drug discovery. Process Biochem, 78: 148–160.

Fong EL, Harrington DA, Farach-Carson MC, et al., 2016, Heralding a new paradigm in 3D tumor modeling. Biomaterials, 108: 197–213.

Zhang C, Yang Z, Dong DL, et al., 2020, 3D culture technologies of cancer stem cells: promising ex vivo tumor models. J Tissue Eng, 11: 1–17.

Shapira A, Dvir T, 2021, 3D tissue and organ printing-hope and reality. Adv Sci, 8(10): 2003751.

Ng WL, Chua CK, Shen Y-F, 2019, Print me an organ! Why we are not there yet. Prog Polym Sci, 97: 101145.

Ma L, Li Y, Wu Y, et al., 2020, The construction of in vitro tumor models based on 3D bioprinting. Bio-Des Manuf, 3(3): 227–236.

Mao S, Pang Y, Liu T, et al., 2020, Bioprinting of in vitro tumor models for personalized cancer treatment: a review. Biofabrication, 12(4): 042001.

Xie F, Sun L, Pang Y, et al., 2021, Three-dimensional bio-printing of primary human hepatocellular carcinoma for personalized medicine. Biomaterials, 265: 120416.

Bahcecioglu G, Basara G, Ellis BW, et al., 2020, Breast cancer models: Engineering the tumor microenvironment. Acta Biomater, 106: 1–21.

Jiang T, Munguia-Lopez JG, Flores-Torres S, et al., 2019, Extrusion bioprinting of soft materials: An emerging technique for biological model fabrication. Appl Phys Rev, 6(1): 011310.

Li X, Liu B, Pei B, et al., 2020, Inkjet bioprinting of biomaterials. Chem Rev, 120(19): 10793–10833.

Ng WL, Lee JM, Zhou M, et al., 2020, Vat polymerization-based bioprinting-process, materials, applications and regulatory challenges. Biofabrication, 12(2): 022001.

Oztan YC, Nawafleh N, Zhou Y, et al., 2020, Recent advances on utilization of bioprinting for tumor modeling. Bioprinting, 18: e00079.

Mazza G, Telese A, Al-Akkad W, et al., 2019, Cirrhotic human liver extracellular matrix 3d scaffolds promote smad-dependent tgf-beta1 epithelial mesenchymal transition. Cells, 9(1): 83.

Hoshiba T, 2019, Decellularized extracellular matrix for cancer research. Materials (Basel), 12(8): 1311.

Kabirian F, Mozafari M, 2020, Decellularized ECM-derived bioinks: Prospects for the future. Methods, 171: 108–118.

Su J, Satchell SC, Wertheim JA, et al., 2019, Poly(ethylene glycol)-crosslinked gelatin hydrogel substrates with conjugated bioactive peptides influence endothelial cell behavior. Biomaterials, 201: 99–112.

Estrada MF, Rebelo SP, Davies EJ, et al., 2016, Modelling the tumour microenvironment in long-term microencapsulated 3D co-cultures recapitulates phenotypic features of disease progression. Biomaterials, 78: 50–61.

Xu K, Wang Z, Copland JA, et al., 2020, 3D porous chitosan-chondroitin sulfate scaffolds promote epithelial to mesenchymal transition in prostate cancer cells. Biomaterials, 254: 120126.

Choi S, Friedrichs J, Song YH, et al., 2019, Intrafibrillar, bone-mimetic collagen mineralization regulates breast cancer cell adhesion and migration. Biomaterials, 198: 95–106.

Alabi BR, Laranger R, Shay JW, 2019, Decellularized mice colons as models to study the contribution of the extracellular matrix to cell behavior and colon cancer progression. Acta Biomater, 100: 213–222.

Suo A, Xu W, Wang Y, et al., 2019, Dual-degradable and injectable hyaluronic acid hydrogel mimicking extracellular matrix for 3D culture of breast cancer MCF-7 cells. Carbohydr Polym, 211: 336–348.

Zhang T, Zhang Q, Chen J, et al., 2014, The controllable preparation of porous PLGA microspheres by the oil/ water emulsion method and its application in 3D culture of ovarian cancer cells. Colloids Surf Physicochem Eng Aspects, 452: 115–124.

Ariadna G-P, Marc R, Teresa P, et al., 2016, Optimization of poli(ɛ-caprolactone) scaffolds suitable for 3D cancer cell culture. Procedia CIRP, 49: 61–66.

Wang C, Li J, Sinha S, et al., 2019, Mimicking brain tumor-vasculature microanatomical architecture via co-culture of brain tumor and endothelial cells in 3D hydrogels. Biomaterials, 202: 35–44.

Ferreira LP, Gaspar VM, Mano JF, 2020, Decellularized extracellular matrix for bioengineering physiomimetic 3D in vitro tumor models. Trends Biotechnol, 38(12): 1397– 1414.

Zhao L, Huang L, Yu S, et al., 2017, Decellularized tongue tissue as an in vitro model for studying tongue cancer and tongue regeneration. Acta Biomater, 58: 122–135.

Narkhede AA, Shevde LA, Rao SS, 2017, Biomimetic strategies to recapitulate organ specific microenvironments for studying breast cancer metastasis. Int J Cancer, 141(6): 1091–1109.

Lang R, Stern MM, Smith L, et al., 2011, Three-dimensional culture of hepatocytes on porcine liver tissue-derived extracellular matrix. Biomaterials, 32(29): 7042–7052.

Zhao F, Cheng J, Sun M, et al., 2020, Digestion degree is a key factor to regulate the printability of pure tendon decellularized extracellular matrix bio-ink in extrusion-based 3D cell printing. Biofabrication, 12(4): 045011.

Pati F, Jang J, Ha DH, et al., 2014, Printing three-dimensional tissue analogues with decellularized extracellular matrix bioink. Nat Commun, 5: 3935.

Xu J, Fang H, Zheng S, et al., 2021, A biological functional hybrid scaffold based on decellularized extracellular matrix/ gelatin/chitosan with high biocompatibility and antibacterial activity for skin tissue engineering. Int J Biol Macromol, 187: 840–849.

Kim BS, Kwon YW, Kong JS, et al., 2018, 3D cell printing of in vitro stabilized skin model and in vivo pre-vascularized skin patch using tissue-specific extracellular matrix bioink: A step towards advanced skin tissue engineering. Biomaterials, 168: 38–53.

Kim J, Shim IK, Hwang DG, et al., 2019, 3D cell printing of islet-laden pancreatic tissue-derived extracellular matrix bioink constructs for enhancing pancreatic functions. J Mater Chem B, 7(10): 1773–1781.

Xu J, Fang H, Su Y, et al., 2022, A 3D bioprinted decellularized extracellular matrix/gelatin/quaternized chitosan scaffold assembling with poly(ionic liquid)s for skin tissue engineering. Int J Biol Macromol, 220: 1253–1266.

Zhang Y, Yuan B, Zhang Y, et al., 2020, Biomimetic lignin/ poly(ionic liquids) composite hydrogel dressing with excellent mechanical strength, self-healing properties, and reusability. Chem Eng J, 400: 125984.

Du J, Hu X, Su Y, et al., 2022, Gelatin/sodium alginate hydrogel-coated decellularized porcine coronary artery to construct bilayer tissue engineered blood vessels. Int J Biol Macromol, 209: 2070–2083.

Coronado RE, Somaraki-Cormier M, Natesan S, et al., 2018, Decellularization and solubilization of porcine liver for use as a substrate for porcine hepatocyte culture. Cell Transplant, 26(12): 1840–1854.

Abaci A, Guvendiren M, 2020, Designing decellularized extracellular matrix-based bioinks for 3D bioprinting. Adv Healthc Mater, 9(24): e2000734.

Sellaro TL, Ranade A, Faulk DM, et al., 2010, Maintenance of human hepatocyte function in vitro by liver-derived extracellular matrix gels. Tissue Eng Part A, 16(3): 1075– 1082.

Saleh T, Ahmed E, Yu L, et al., 2018, Silver nanoparticles improve structural stability and biocompatibility of decellularized porcine liver. Artif Cells Nanomed Biotechnol, 46(sup2): 273–284.

Wu Q, Bao J, Zhou YJ, et al., 2015, Optimizing perfusion-decellularization methods of porcine livers for clinical-scale whole-organ bioengineering. Biomed Res Int, 2015: 785474.

Poornejad N, Nielsen JJ, Morris RJ, et al., 2016, Comparison of four decontamination treatments on porcine renal decellularized extracellular matrix structure, composition, and support of human renal cortical tubular epithelium cells. J Biomater Appl, 30(8): 1154–1167.

Struecker B, Hillebrandt KH, Voitl R, et al., 2015, Porcine liver decellularization under oscillating pressure conditions: A technical refinement to improve the homogeneity of the decellularization process. Tissue Eng Part C Methods, 21(3): 303–313.

Sun D, Liu Y, Wang H, et al., 2018, Novel decellularized liver matrix-alginate hybrid gel beads for the 3D culture of hepatocellular carcinoma cells. Int J Biol Macromol, 109: 1154–1163.

Hu X, Li W, Li L, et al., 2019, A biomimetic cartilage gradient hybrid scaffold for functional tissue engineering of cartilage. Tissue Cell, 58: 84–92.

Li W, Hu X, Yang S, et al., 2018, A novel tissue-engineered 3D tumor model for anti-cancer drug discovery. Biofabrication, 11(1): 015004.

Mao S, He J, Zhao Y, et al., 2020, Bioprinting of patient-derived in vitro intrahepatic cholangiocarcinoma tumor model: Establishment, evaluation and anti-cancer drug testing. Biofabrication, 12(4): 045014.

Song K, Li L, Yan X, et al., 2017, Characterization of human adipose tissue-derived stem cells in vitro culture and in vivo differentiation in a temperature-sensitive chitosan/beta-glycerophosphate/collagen hybrid hydrogel. Mater Sci Eng C Mater Biol Appl, 70(Pt 1): 231–240.

Ijima H, Nakamura S, Bual R, et al., 2018, Physical properties of the extracellular matrix of decellularized porcine liver. Gels, 4(2): 39.

Polley C, Mau R, Lieberwirth C, et al., 2017, Bioprinting of three dimensional tumor models: a preliminary study using a low cost 3D printer. Curr Dir Biomed Eng, 3(2): 135–138.

Lee H, Han W, Kim H, et al., 2017, Development of liver decellularized extracellular matrix bioink for three-dimensional cell printing-based liver tissue engineering. Biomacromolecules, 18(4): 1229–1237.

Chaji S, Al-Saleh J, Gomillion CT, 2020, Bioprinted three-dimensional cell-laden hydrogels to evaluate adipocyte-breast cancer cell interactions. Gels, 6(1): 10.

Lv K, Zhu J, Zheng S, et al., 2021, Evaluation of inhibitory effects of geniposide on a tumor model of human breast cancer based on 3D printed Cs/Gel hybrid scaffold. Mater Sci Eng C, 119: 111509.

Jeong W, Kim MK, Kang HW, 2021, Effect of detergent type on the performance of liver decellularized extracellular matrix-based bio-inks. J Tissue Eng, 12: 1–14.

Bordoni M, Karabulut E, Kuzmenko V, et al., 2020, 3D printed conductive nanocellulose scaffolds for the differentiation of human neuroblastoma cells. Cells, 9(3): 682.

Zhao C, Li Y, Peng G, et al., 2020, Decellularized liver matrix-modified chitosan fibrous scaffold as a substrate for C3A hepatocyte culture. J Biomater Sci, Polym Ed, 31(8): 1041–1056.

Lu S, Cuzzucoli F, Jiang J, et al., 2018, Development of a biomimetic liver tumor-on-a-chip model based on decellularized liver matrix for toxicity testing. Lab Chip, 18(22): 3379–3392.

Wang JZ, Zhu YX, Ma HC, et al., 2016, Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/ collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. Mater Sci Eng C Mater Biol Appl, 62: 215–225.

Sachlos E, Reis N, Ainsley C, et al., 2003, Novel collagen scaffolds with predefined internal morphology made by solid freeform fabrication. Biomaterials, 24(8): 1487–1497.

Xu W, Qian J, Zhang Y, et al., 2016, A double-network poly(Nvarepsilon-acryloyl L-lysine)/hyaluronic acid hydrogel as a mimic of the breast tumor microenvironment. Acta Biomater, 33: 131–141.

Li X, Deng Q, Zhuang T, et al., 2020, 3D bioprinted breast tumor model for structure–activity relationship study. Bio- Des Manuf, 3(4): 361–372.

Peela N, Sam FS, Christenson W, et al., 2016, A three dimensional micropatterned tumor model for breast cancer cell migration studies. Biomaterials, 81: 72–83.

Antunes J, Gaspar VM, Ferreira L, et al., 2019, In-air production of 3D co-culture tumor spheroid hydrogels for expedited drug screening. Acta Biomater, 94: 392–409.

Li L, Qin S, Peng J, et al., 2020, Engineering gelatin-based alginate/carbon nanotubes blend bioink for direct 3D printing of vessel constructs. Int J Biol Macromol, 145: 262–271.

Xu K, Ganapathy K, Andl T, et al., 2019, 3D porous chitosan-alginate scaffold stiffness promotes differential responses in prostate cancer cell lines. Biomaterials, 217: 119311.

Mahmoudzadeh A, Mohammadpour H, 2016, Tumor cell culture on collagen-chitosan scaffolds as three-dimensional tumor model: A suitable model for tumor studies. J Food Drug Anal, 24(3): 620–626.

Ananthanarayanan B, Kim Y, Kumar S, 2011, Elucidating the mechanobiology of malignant brain tumors using a brain matrix-mimetic hyaluronic acid hydrogel platform. Biomaterials, 32(31): 7913–7923.

Kwak BS, Choi W, Jeon J-W, et al., 2018, In vitro 3D skin model using gelatin methacrylate hydrogel. J Ind Eng Chem, 66: 254–261.

Pulaski BA, S. O-R, 2000, Mouse 4T1 breast tumor model. Curr Protoc Immunol, 39(1): 1–16.

Gregorio AC, Fonseca NA, Moura V, et al., 2016, Inoculated cell density as a determinant factor of the growth dynamics and metastatic efficiency of a breast cancer murine model. PLoS One, 11(11): e0165817.

Keklikoglou I, Cianciaruso C, Guc E, et al., 2019, Chemotherapy elicits pro-metastatic extracellular vesicles in breast cancer models. Nat Cell Biol, 21(2): 190–202.

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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing