Miniaturized Flow-Through Bioreactor for Processing and Testing in Pharmacology

Abstract:

Conventional Bioreactor systems for cultivating cells in Life Science have been widely used for decades. An in vitro cell cultivation bioreactor should reliably and reproducibly mimic the in vivo microenvironment of the cultured cells. Normally, mammalian cell cultures are performed in conventional bioreactor devices such as culture flasks and culture-dishes. However, these tools have fundamental limitations due to being inappropriate for high throughput screening and consume a considerable amount of resources and time [1]. Therefore, there is a trend towards miniaturization, disposables and even micro platforms that fulfill increasing demands strongly aiming for production and testing of novel pharmaceutical products. Here we present the development and manufacture of a disposable miniaturized flow-through bioreactor system that can be produced in large numbers at low costs. nanoporous hollow fibers are located at the fluidic sources and drains of the miniaturized bioreactors and retain cells. The necessary mixture of oxygen and carbon dioxide is provided via diffusion through a semi-permeable membrane. Fluidic connections allow the continuous feeding of the cells adding nutrient solution at constant rates at the inlet of the micro bioreactor and removing the solution at the same rate at the outlet. This medium can be collected and used for subsequent analysis. Different designs and concepts for such bioreactors were carried out with varying numbers of plates, and integrated or joined miniaturized reactor chambers. First tests show full technical and biological functionality, cells could successfully be cultivated at high viability rates for some days.

• Periodical: Materials Science Forum (Volume 879)
• Pages: 236-243
• DOI: https://doi.org/10.4028/www.scientific.net/MSF.879.236
• Online since: November 2016
• Authors: Andrea Böhme*, Lars Radke, Felix Schütze, Sylvio Schneider, Thilo Liebscher, Sabine Sauer, Loredana Santo, Fabrizio Quadrini, Michael Hummel, Christoph Giese, Marcus Frohme, Andreas H. Foitzik
• Keywords: Bioreactor, Miniaturization, Perfusion, Pharmacology, Tightness Testing

References

[1] A. Abbott, Biology's new dimension, Nature, 424 (6951) (2003), 870-872.

[2] D.B. Weibel, G.M. Whitesides, Applications of microfluidics in chemical biology, Curr. Opin. Chem. Biol. 10 (6) (2006), 584-591.

[3] C. Giese, U. Marx, Human immunity in vitro — Solving immunogenicity and more, Adv Drug Deliv Rev. 69–70 (2014) 103–122, doi: 10. 1016/j. addr. 2013. 12. 011.

DOI: 10.1016/j.addr.2013.12.011

[4] M. Leist, T. Hartung, Inflammatory findings on species extrapolations: humans are definitely no 70-kg mice, Arch. Toxicol. 87 (2013) 563–567.

DOI: 10.1007/s00204-013-1038-0

[5] G. Suntharalingam, M.R. Perry, S. Ward, S.J. Brett, A. Castello-Cortes, M.D. Brunner, N. Panoskaltsis, Cytokine storm in a phase 1 trial of the anti-CD28 monoclonal antibody TGN1412, N Engl J Med 355 (2006) 1018–1028.

DOI: 10.1056/nejmoa063842

[6] W.L. Warren, D.D. Iii, J. Moser, S. Inderpal, H. Song, E. Mishkin; J.G. Tew, Co-culture lymphoid tissue equivalent (LTE) for an artificial immune system (AIS), Pat. WO2007146334 A2, (2007).

[7] L.G. Griffith, M.A. Swartz , Capturing complex 3D tissue physiology in vitro, Nat Rev Mol Cell Biol 7(3) (2006) 211-24.

DOI: 10.1038/nrm1858

[8] C. Dieterich, S. Rupp, M. Noll, T. Graeve, In Vitro-Gewebetestsystem - Cartilage replacement, useful particularly for repairing joint defects, comprises differentiated and proliferating chondrocytes in collagen-based biomatrix. Pat. DE10062626 (A1), (2001).

[9] C. Giese, C.D. Demmler, R. Ammer, S. Hartmann, A. Lubitz, L. Miller, R. Müller, U. Marx, A human lymph node in vitro—challenges and progress, Artif. Organs 30 (2006) 803–808.

DOI: 10.1111/j.1525-1594.2006.00303.x

[10] M. Birkholz, K. -E. Ehwald, T. Basmer, P. Kulse, C. Reich, J. Drews, D. Genschow, U. Haak, S. Marschmeyer, E. Matthus, K. Schulz, D. Wolansky, W. Winkler, T. Guschauski, R. Ehwald, Sensing glucose concentrations at GHz frequencies with a fully embedded Biomicro-electromechanical system (BioMEMS), J Appl Phys 113 (24): 244904 (2013).

DOI: 10.1063/1.4811351

[10] E.M. Materne, A.G. Tonevitsky, U. Marx, Chip-based liver equivalents for toxicity testing — organotypicalness versus cost-efficient high throughput, Lab Chip 13 (2013) 3481–3495.

DOI: 10.1039/c3lc50240f

[11] M. Noll, T. Graeve, Three-Dimensional Skin Model, Pat. EP 1 290 145 B1, (2001).

[12] M. Seifert, A. Lubitz, J. Trommer, D. Könnig, G. Korus, U. Marx, H. Volk, G. Duda, G. Kasper, K. Lehmann, M. Stolk, C. Giese, Crosstalk between immune cells and mesenchymal stromal cells in a 3D bioreactor system, Int. J. Artif. Organs 35 (2012).

DOI: 10.5301/ijao.5000118

[13] K. Schulze, D.A. López, U.M. Tillich, M. Frohme, A simple viability analysis for unicellular cyanobacteria using a new autofluorescence assay, automated microscopy, and ImageJ. BMC Biotechnol. 11: 118 (2011), DOI: 10. 1186/1472-6750-11-118.

DOI: 10.1186/1472-6750-11-118

[14] L. Radke, D.A. López, A. Lubitz, C. Giese, M. Frohme, Induced cytokine response of human PMBC-cultures: Correlation of gene expression and secretion profiling and the effect of cryopreservation, Cell. Immunol. 272(2) (2012) 144–153.

DOI: 10.1016/j.cellimm.2011.10.018

[15] C. Giese, A. Lubitz, C.D. Demmler, J. Reuschel, K. Bergner, U. Marx, Immunological substance testing on human lymphatic micro-organoids in vitro, J. Biotechnol. 148 (2010) 38–45.

DOI: 10.1016/j.jbiotec.2010.03.001