Abstract
The creation of larger-scale three-dimensional tissue constructs depends on proper medium mass and gas exchange, as well as removal of metabolites, which cannot be achieved in conventional static two-dimensional petri dish culture. In cultures of tissue-density this problem can be addressed by decentral perfusion through artificial micro-capillaries. While the static medium exchange in petri dishes leads to metabolite peaks, perfusion culture provides a dynamic medium supply, thereby preventing non-physiological peaks. To overcome the limitations of conventional static two-dimensional culture, a three-dimensional perfusion bioreactor technology has been developed, providing decentral and high-performance mass exchange as well as integral oxygenation. Similar to organ systems in vivo, the perfusion with medium provides nutrition and removes waste metabolites, and the perfusion with gas delivers oxygen and carbon dioxide for pH regulation. Such bioreactors are available at various dimensions ranging from 0.2 to 800 mL cell compartment volumes (manufactured by StemCell Systems, Berlin, Germany). Here, we describe in detail the setup and maintenance of a small-scale 4-chamber bioreactor with its tubing circuit and perfusion system.
Keywords:
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Pittman RN (2011) The circulatory system and oxygen transport. In: Regulation of tissue oxygenation. Morgan & Claypool Life Sciences, San Rafael, CA. doi: 10.4199/C00029ED1V01Y201103ISP017
Busse B, Smith MD, Gerlach JC (1999) Treatment of acute liver failure: hybrid liver support. A critical overview. Langenbecks Arch Surg 384(6):588–599
Sauer I, Zeilinger K, Pless G, Kardassis D, Theruvath T, Pascher A, Mueller A, Steinmueller T, Neuhaus P, Gerlach J (2003) Extracorporeal liver support based on primary human liver cells and albumin dialysis—treatment of a patient with primary graft nonfunction. J Hepatol 39(4):649–653
Sauer IM, Kardassis D, Zeillinger K, Pascher A, Gruenwald A, Pless G, Irgang M, Kraemer M, Puhl G, Frank J, Muller AR, Steinmuller T, Denner J, Neuhaus P, Gerlach JC (2003) Clinical extracorporeal hybrid liver support-phase I study with primary porcine liver cells. Xenotransplantation 10:460–469
Gerlach JC, Brayfield C, Puhl G, Borneman R, Muller C, Schmelzer E, Zeilinger K (2010) Lidocaine/monoethylglycinexylidide test, galactose elimination test, and sorbitol elimination test for metabolic assessment of liver cell bioreactors. Artif Organs 34(6):462–472, doi: AOR885 [pii] 10.1111/j.1525-1594.2009.00885.x
Gerlach JC, Hout M, Edsbagge J, Bjorquist P, Lubberstedt M, Miki T, Stachelscheid H, Schmelzer E, Schatten G, Zeilinger K (2009) Dynamic 3D culture promotes spontaneous embryonic stem cell differentiation in vitro. Tissue Eng Part C Methods 16(1):115–121, doi: 10.1089/ten.TEC.2008.0654
Gerlach JC, Lubberstedt M, Edsbagge J, Ring A, Hout M, Baun M, Rossberg I, Knospel F, Peters G, Eckert K, Wulf-Goldenberg A, Bjorquist P, Stachelscheid H, Urbaniak T, Schatten G, Miki T, Schmelzer E, Zeilinger K (2010) Interwoven four-compartment capillary membrane technology for three-dimensional perfusion with decentralized mass exchange to scale up embryonic stem cell culture. Cells Tissues Organs 192(1):39–49, doi: 000291014 [pii] 10.1159/000291014
Gerlach JC, Over P, Foka HG, Turner ME, Thompson RL, Gridelli B, Schmelzer E (2014) Role of transcription factor CCAAT/enhancer-binding protein alpha in human fetal liver cell types in vitro. Hepatol Res 45(8):919–932. doi:10.1111/hepr.12420
Pekor C, Gerlach JC, Nettleship I, Schmelzer E (2015) Induction of hepatic and endothelial differentiation by perfusion in a three-dimensional cell culture model of human fetal liver. Tissue Eng Part C Methods. doi:10.1089/ten.TEC.2014.0453
Schmelzer E, Mutig K, Schrade P, Bachmann S, Gerlach JC, Zeilinger K (2009) Effect of human patient plasma ex vivo treatment on gene expression and progenitor cell activation of primary human liver cells in multi-compartment 3D perfusion bioreactors for extra-corporeal liver support. Biotechnol Bioeng 103(4):817–827. doi:10.1002/bit.22283
Schmelzer E, Triolo F, Turner ME, Thompson RL, Zeilinger K, Reid LM, Gridelli B, Gerlach JC (2010) Three-dimensional perfusion bioreactor culture supports differentiation of human fetal liver cells. Tissue Eng Part A 16(6):2007–2016. doi:10.1089/ten.TEA.2009.0569
Stachelscheid H, Wulf-Goldenberg A, Eckert K, Jensen J, Edsbagge J, Bjorquist P, Rivero M, Strehl R, Jozefczuk J, Prigione A, Adjaye J, Urbaniak T, Bussmann P, Zeilinger K, Gerlach JC (2013) Teratoma formation of human embryonic stem cells in three-dimensional perfusion culture bioreactors. J Tissue Eng Regen Med 7(9):729–741. doi:10.1002/term.1467
Hoffmann SA, Muller-Vieira U, Biemel K, Knobeloch D, Heydel S, Lubberstedt M, Nussler AK, Andersson TB, Gerlach JC, Zeilinger K (2012) Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies. Biotechnol Bioeng 109(12):3172–3181. doi:10.1002/bit.24573
Mueller D, Tascher G, Muller-Vieira U, Knobeloch D, Nuessler AK, Zeilinger K, Heinzle E, Noor F (2011) In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor. J Tissue Eng Regen Med 5(8):e207–e218. doi:10.1002/term.418
Housler GJ, Miki T, Schmelzer E, Pekor C, Zhang X, Kang L, Voskinarian-Berse V, Abbot S, Zeilinger K, Gerlach JC (2012) Compartmental hollow fiber capillary membrane-based bioreactor technology for in vitro studies on red blood cell lineage direction of hematopoietic stem cells. Tissue Eng Part C Methods 18(2):133–142. doi:10.1089/ten.TEC.2011.0305
Housler GJ, Pekor C, Miki T, Schmelzer E, Zeilinger K, Gerlach JC (2014) 3-D perfusion bioreactor process optimization for CD34+ hematopoietic stem cell culture and differentiation towards red blood cell lineage. J Bone Marrow Res 2:3. doi:10.4172/2329-8820.1000150
Darnell M, Schreiter T, Zeilinger K, Urbaniak T, Soderdahl T, Rossberg I, Dillner B, Berg AL, Gerlach JC, Andersson TB (2011) Cytochrome P450-dependent metabolism in HepaRG cells cultured in a dynamic three-dimensional bioreactor. Drug Metab Dispos 39(7):1131–1138. doi:10.1124/dmd.110.037721
Miki T, Ring A, Gerlach J (2011) Hepatic differentiation of human embryonic stem cells is promoted by three-dimensional dynamic perfusion culture conditions. Tissue Eng Part C Methods 17(5):557–568. doi:10.1089/ten.TEC.2010.0437
Ring A, Gerlach J, Peters G, Pazin BJ, Minervini CF, Turner ME, Thompson RL, Triolo F, Gridelli B, Miki T (2010) Hepatic maturation of human fetal hepatocytes in four-compartment three-dimensional perfusion culture. Tissue Eng Part C Methods 16(5):835–845. doi:10.1089/ten.TEC.2009.0342
Zeilinger K, Schreiter T, Darnell M, Soderdahl T, Lubberstedt M, Dillner B, Knobeloch D, Nussler AK, Gerlach JC, Andersson TB (2011) Scaling down of a clinical three-dimensional perfusion multicompartment hollow fiber liver bioreactor developed for extracorporeal liver support to an analytical scale device useful for hepatic pharmacological in vitro studies. Tissue Eng Part C Methods 17(5):549–556. doi:10.1089/ten.TEC.2010.0580
Gerlach JC, Encke J, Hole O, Muller C, Ryan CJ, Neuhaus P (1994) Bioreactor for a larger scale hepatocyte in vitro perfusion. Transplantation 58(9):984–988
Gerlach J, Schnoy N, Encke J, Müller C, Smith M, Neuhaus P (1995) Improved hepatocyte in vitro maintenance in a culture model with woven multicompartment capillary systems: electron microscopy studies. Hepatology 22:546–552
Gerlach JC, Witaschek T, Strobel C, Brayfield CA, Bornemann R, Catapano G, Zeilinger K (2010) Feasibility of using sodium chloride as a tracer for the characterization of the distribution of matter in complex multi-compartment 3D bioreactors for stem cell culture. Int J Artif Organs 33(6):399–404
Balmert SC, McKeel D, Triolo F, Gridelli B, Zeilinger K, Bornemann R, Gerlach JC (2011) Perfusion circuit concepts for hollow-fiber bioreactors used as in vitro cell production systems or ex vivo bioartificial organs. Int J Artif Organs 34(5):410–421. doi:10.5301/ijao.2011.8366
Sussman NL, Kelly JH (1993) Improved liver function following treatment with an extracorporeal liver assist device. Artif Organs 17(1):27–30
Gerlach J, Neuhaus P (1994) Culture model for primary hepatocytes. In Vitro Cell Dev Biol 30A:640–642
Acknowledgements
This work was supported by NIH grant 5R01HL108631 and by the University of Pittsburgh Medical Center. The authors thank Daniel McKeel for his continuous excellent technical support.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Schmelzer, E., Gerlach, J.C. (2016). Multicompartmental Hollow-Fiber-Based Bioreactors for Dynamic Three-Dimensional Perfusion Culture. In: Turksen, K. (eds) Bioreactors in Stem Cell Biology. Methods in Molecular Biology, vol 1502. Humana Press, New York, NY. https://doi.org/10.1007/7651_2016_335
Download citation
DOI: https://doi.org/10.1007/7651_2016_335
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6476-5
Online ISBN: 978-1-4939-6478-9
eBook Packages: Springer Protocols