Validation of a novel perfusion bioreactor system in cancer research
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Abstract
Development of drugs is a complex, time- and cost-consuming process due to the lack of standardized and reliable characterization techniques and models. Traditionally, drug screening is based on in vitro analysis using two-dimensional (2D) cell cultures followed by in vivo animal testing. Unfortunately, application of the obtained results to humans in about 90 % of cases fails. Therefore, it is important to develop and improve cell-based systems that can mimic the in vivo- like conditions to provide more reliable results. In this paper, we present development and validation of a novel user-friendly perfusion bioreactor system for single use aimed for cancer research, drug screening, anti-cancer drug response studies, biomaterial characterization and tissue engineering. Simple design of the perfusion bioreactor provides direct medium flow at physiological velocities (100–250 µm s-1) through samples of different sizes and shapes. Biocompatibility of the bioreactor was confirmed in short term cultivation studies of cervical carcinoma SiHa cells immobilized in alginate microfibers under continuous medium flow. The results have shown preserved cell viability indicating that the perfusion bioreactor in conjunction with alginate hydrogels as cell carriers could be potentially used as a tool for controlled anti-cancer drug screening in a 3D environment.
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Luo Y, Wang C, Hussein M, Qiao Y, Ma L, An J, Su M. Three-dimensional microtissue assay for high-throughput cytotoxicity of nanoparticles. Anal Chem. 2012; 84: 6731−6738.
Li Q, Lin H, Rauch J, Deleyrolle L, Reynolds B, Viljoen H, Zhang C, Zhang C, Gu L, Van Wyk E, Lei Y. Scalable culturing of primary human glioblastoma tumor- initiating cells with a cell-friendly culture system. Sci Rep. 2018; 8(3531): 1-12.
Mazzoleni G, Di Lorenzo D, Steimberg N. Modelling tissues in 3D: the next future of pharmaco-toxicology and food research? Genes Nutr. 2009; 4(1): 13-22.
Edmondson R, Broglie JJ, Adcock A, Yang L. Three dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol. 2014; 12: 207-218.
Akay M, Hite J, Avci NG, Fan Y, Akay Y, Lu G, Zhu J-J. Drug screening of human GBM spheroids in brain cancer chip. Sci Rep. 2018; 8 (15423): 1-9.
Joris F, Manshian BB, Peynshaert K, De Smedt SC, Braeckmansad K, Soenen SJ. Assessing nanoparticle toxicity in cell-based assays: Influence of cell culture parameters and optimized models for bridging the in vitro-in vivo gap. Chem Soc Rev. 2013; 42(21): 8339-8359.
Mak IW, Evaniew N, Ghert M. Lost in translation: animal models and clinical trials in cancer treatment. Am J Transl Res. 2014; 6(2): 114-118.
Akhtar A. The flaws and human harms of animal experimentation. Camb Q Healthc Ethics. 2015; 24(4): 407–419.
Osmokrovic A, Obradovic B, Bugarski D, Bugarski B, Vunjak-Novakovic G. Development of a packed bed bioreactor for cartilage tissue engineering. FME Trans. 2006; 34: 65–70.
Grayson WL, Bhumiratana S, Cannizzaro C, Chao PH, Lennon DP, Caplan AI, Vunjak-Novakovic G. Effects of initial seeding density and fluid perfusion rate on formation of tissue-engineered bone. Tissue Eng Part A. 2008; 14: 1809–1820.
Ravichandran A, Wen F, Lim J, Chong MSK, Chan JKY, Teoh SH. Biomimetic fetal rotation bioreactor for engineering bone tissues—Effect of cyclic strains on upregulation of osteogenic gene expression. J Tissue Eng Regen Med. 2018; 12(4): e2039-e2050.
Zvicer J, Miskovic-Stankovic V, Obradovic B. Functional bioreactor characterization to assess potentials of nanocomposites based on different alginate types and silver nanoparticles for use as cartilage tissue implants. J Biomed Mater Res A. 2019; 107: 755-768.
Costa E, Moreira A, de Melo-Diogo D, Gaspar V, Carvalho M, Correia I. 3D tumor spheroids: an overview on the tools and techniques used for their analysis. Biotechnol Adv. 2016; 34: 1427-1441.
Lazzari G, Nicolas V, Matsusaki M, Akashi M, Couvreur P, Mura S. Multicellular spheroid based on a triple co-culture: A novel 3D model to mimic pancreatic tumor complexity. Acta Biomater. 2018; 78: 296–307.
Friedrich J, Seidel C, Ebner R, Kunz-Schughart LA. Spheroid-based drug screen: considerations and practical approach. Nat Protoc. 2009; 4(3): 309-324.
Wan X, Li Z , Ye H, Cui Z. Three-dimensional perfused tumour spheroid model for anti-cancer drug screening. Biotechnol Lett 2016; 38:1389–1395.
Stojkovska J, Djurdjevic Z, Jancic I, Bufan B, Milenkovic M, Jankovic R, Miskovic-Stankovic V, Obradovic B. Comparative in vivo evaluation of novel formulations based on alginate and silver nanoparticles for wound treatments. J Biomater Appl. 2018; 32: 1197–1211.
Stojkovska J, Bugarski B, Obradovic B. Evaluation of alginate hydrogels under in vivo–like bioreactor conditions for cartilage tissue engineering. J Mater Sci Mater Med. 2010; 21: 2869–2879.
Cacciotti I, Ceci C, Bianco A, Pistritto G. Neuro-differentiated Ntera2 cancer stem cells encapsulated in alginate beads: First evidence of biological functionality. Mater Sci Eng C. 2017; 81: 32-38.
Obradović B, Stojkovska J, Zvicer J. Single-use perfusion chamber for cell and tissue culture and biomaterial characterization, Patent application RS P-2018/0569, 2018.
Briers JD, Webster S. Laser speckle contrast analysis (LASCA): a nonscanning, full-field technique for monitoring capillary blood flow. J Biomed Opt. 1996; 1(2): 174–179.
Galateanu B, Dimonie D, Vasile E, Nae S, Cimpean A, Costache M. Layer-shaped alginate hydrogels enhance the biological performance of human adipose-derived stem cells. BMC Biotechnol. 2012;12:35.
Wang Q, Li S, Xie Y, Yu W, Xiong Y, Ma X, Yuan Q. Cytoskeletal reorganization and repolarization of hepatocarcinoma cells in APA microcapsule to mimic native tumor characteristics. Hepatol Res. 2006; 35(2): 96-103.
Garland EM, Parr JM, Williamson DS, Cohen SM. In vitro cytotoxicity of the sodium, potassium and calcium salts of saccharin, sodium ascorbate, sodium citrate and sodium chloride. Toxicol In Vitro. 1989; 3(3): 201-205.
Lu Y, Zhang X, Zhang H, Lan J, Huang G, Varin E, Lincet H, Poulain L, Icard P. Citrate induces apoptotic cell death: a promising way to treat gastric carcinoma? Anticancer Res. 2011; 31(3): 797-806.
Xia Y, Zhang X, Bo A, Sun J, Li M. Sodium citrate inhibits the proliferation of human gastric adenocarcinoma epithelia cells. Oncol Lett. 2018; 15: 6622-6628.
CaiazzaC, DAgostino M, Passaro F, Faicchia D, Mallardo M, Paladino S, Pierantoni GM, Tramontano D. Effects of long-term citrate treatment in the PC3 prostate cancer cell line. Int J Mol Sci. 2019; 20(11): 2613.
Lee GM, Gray JJ, Palsson B. Effect of trisodium citrate treatment on hybridoma cell viability. Biotechnol Tech. 1991; 5: 295–298.
Hammer J, Han LH, Tong X, Yang F. A facile method to fabricate hydrogels with microchannel-like porosity for tissue engineering. Tissue Eng Part C Methods. 2014; 20(2): 169-176.
Chen CY, Ke CJ, Yen KC, Hsieh HC, Sun JS, Lin FH. 3D porous calcium-alginate scaffolds cell culture system improved human osteoblast cell clusters for cell therapy. Theranostics. 2015 5(6):643-55.