Abstract
Given the increased recognition of the importance of physiologically relevant microenvironments when designing in vitro assays, microphysiological systems (MPS) that mimic the critical function and structure of tissues and organs have gained considerable attention as alternatives to traditional experimental models. Accordingly, the field is growing rapidly, and some promising MPS are being tested for use in pharmaceutical development and toxicological testing. However, most MPS are complex and require additional infrastructure, which limits their successful translation. Here, we present a pumpless, modular MPS consisting of 1) a resistance module that controls flow rate and 2) a physiologically relevant, three-dimensional blood vessel module. Flow is provided by an attached reservoir tank that feeds fluid into the resistance channel via hydrostatic pressure. The flow rate is controlled by the height of the media in the tank and the resistance channel’s dimensions. The flow from the resistance module is streamed into the blood vessel module using a liquid bridge. We utilize optical coherence tomography (OCT) to measure fluid velocity at regions of interest. The endothelial cells cultured in the MPS remain viable for up to 14 days and demonstrate the functional characteristics of the human blood vessels verified by tight junction expression and diffusion assay. Our results show that a modular MPS can simulate a functional endothelium in vitro while simplifying the operation of the MPS. The simplicity of the system allows for modifications to incorporate other microenvironmental components and to build other organ-modeling systems easily.
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S. Alimperti, T. Mirabella, V. Bajaj, W. Polacheck, D.M. Pirone, J. Duffield, C.S. Chen, Three-dimensional biomimetic vascular model reveals a RhoA, Rac1, and N-cadherin balance in mural cell-endothelial cell-regulated barrier function. Proc. Natl. Acad. Sci. U S A 114(33), 8758–8763 (2017). https://doi.org/10.1073/pnas.1618333114
J. Atencia, D.J. Beebe, Steady flow generation in microcirculatory systems. Lab Chip 6(4), 567–574 (2006). https://doi.org/10.1039/b514070f
J.-P. Barral, A. Croibier, General organization of the cardiovascular system, in Visceral Vascular Manipulations. ed. by J.-P. Barral, A. Croibier (Churchill Livingstone, Edinburgh, 2011), pp. 3–26
E. Berthier, D.J. Beebe, Flow rate analysis of a surface tension driven passive micropump. Lab Chip 7(11), 1475–1478 (2007). https://doi.org/10.1039/b707637a
L.L. Bischel, K.E. Sung, J.A. Jimenez-Torres, B. Mader, P.J. Keely, D.J. Beebe, The importance of being a lumen. FASEB J 28(11), 4583–4590 (2014). https://doi.org/10.1096/fj.13-243733
C.F. Buchanan, S.S. Verbridge, P.P. Vlachos, M.N. Rylander, Flow shear stress regulates endothelial barrier function and expression of angiogenic factors in a 3D microfluidic tumor vascular model. Cell Adh. Migr. 8(5), 517–524 (2014). https://doi.org/10.4161/19336918.2014.970001
F. Bunge, S.V.D. Driesche, M.J. Vellekoop, Microfluidic platform for the long-term on-chip cultivation of mammalian cells for lab-on-a-chip applications. Sensors (Basel) 17(7) (2017). https://doi.org/10.3390/s17071603
S.-L. Chen, Z. Xie, P.L. Carson, X. Wang, L.J. Guo, In vivo flow speed measurement of capillaries by photoacoustic correlation spectroscopy. Opt. Lett. 36(20), 4017–2019 (2015). https://doi.org/10.1364/OL.36.004017
K.M. Chrobak, D.R. Potter, J. Tien, Formation of perfused, functional microvascular tubes in vitro. Microvasc. Res. 71(3), 185–196 (2006). https://doi.org/10.1016/j.mvr.2006.02.005
B.S. Conklin, R.P. Vito, C. Chen, Effect of low shear stress on permeability and occludin expression in porcine artery endothelial cells. World J. Surg. 31(4), 733–743 (2007). https://doi.org/10.1007/s00268-006-0735-8
P.M. Davidson, J. Sliz, P. Isermann, C. Denais, J. Lammerding, Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments. Integr. Biol. (Camb). 7(12), 1534–1546 (2015). https://doi.org/10.1039/c5ib00200a
K. De Ceunynck, C.G. Peters, A. Jain, S.J. Higgins, O. Aisiku, J.L. Fitch-Tewfik, R. Flaumenhaft, PAR1 agonists stimulate APC-like endothelial cytoprotection and confer resistance to thromboinflammatory injury. Proc. Natl. Acad. Sci. U S A 115(5), E982–E991 (2018). https://doi.org/10.1073/pnas.1718600115
Y.-Q. Fan, H.-L. Wang, K.-X. Gao, J.-J. Liu, D.-P. Chai, Y.-J. Zhang, Applications of modular microfluidics technology. Chin. J. Anal. Chem. 46(12), 1863–1871 (2018). https://doi.org/10.1016/s1872-2040(18)61126-0
C. Franco, H. Gerhardt, Blood vessels on a chip. Nature 488(7412), 465–466 (2012). https://doi.org/10.1038/488465a
K. Gold, A.K. Gaharwar, A. Jain, Emerging trends in multiscale modeling of vascular pathophysiology: Organ-on-a-chip and 3D printing. Biomaterials 196, 2–17 (2019). https://doi.org/10.1016/j.biomaterials.2018.07.029
M.M. Gong, K.M. Lugo-Cintron, B.R. White, S.C. Kerr, P.M. Harari, D.J. Beebe, Human organotypic lymphatic vessel model elucidates microenvironment-dependent signaling and barrier function. Biomaterials 214, 119225 (2019). https://doi.org/10.1016/j.biomaterials.2019.119225
K.M. Gray, K.M. Stroka, Vascular endothelial cell mechanosensing: New insights gained from biomimetic microfluidic models. Semin. Cell Dev. Biol. 71, 106–117 (2017). https://doi.org/10.1016/j.semcdb.2017.06.002
R.D. Hamilton, A.J. Foss, L. Leach, Establishment of a human in vitro model of the outer blood-retinal barrier. J. Anat. 211(6), 707–716 (2007). https://doi.org/10.1111/j.1469-7580.2007.00812.x
R. Hernandez Vera, P. O’Callaghan, N. Fatsis-Kavalopoulos, J. Kreuger, Modular microfluidic systems cast from 3D-printed molds for imaging leukocyte adherence to differentially treated endothelial cultures. Sci. Rep. 9(1), 11321 (2019). https://doi.org/10.1038/s41598-019-47475-z
A. Jain, R. Barrile, A.D. van der Meer, A. Mammoto, T. Mammoto, K. De Ceunynck, D.E. Ingber, Primary human lung alveolus-on-a-chip model of intravascular thrombosis for assessment of therapeutics. Clin. Pharmacol. Ther. 103(2), 332–340 (2018). https://doi.org/10.1002/cpt.742
E. Jastrzebska, A. Zuchowska, S. Flis, P. Sokolowska, M. Bulka, A. Dybko, Z. Brzozka, Biological characterization of the modified poly(dimethylsiloxane) surfaces based on cell attachment and toxicity assays. Biomicrofluidics 12(4), 044105 (2018). https://doi.org/10.1063/1.5035176
Q. Ji, J.M. Zhang, Y. Liu, X. Li, P. Lv, D. Jin, H. Duan, A Modular microfluidic device via multimaterial 3D printing for emulsion generation. Sci. Rep. 8(1), 4791 (2018). https://doi.org/10.1038/s41598-018-22756-1
J.A. Jimenez-Torres, S.L. Peery, K.E. Sung, D.J. Beebe, LumeNEXT: a practical method to pattern luminal structures in ECM gels. Adv. Healthc. Mater. 5(2), 198–204 (2016). https://doi.org/10.1002/adhm.201500608
H.J. Kim, D. Huh, G. Hamilton, D.E. Ingber, Human gut-on-a-chip inhabited by microbial flora that experiences intestinal peristalsis-like motions and flow. Lab Chip 12(12), 2165–2174 (2012). https://doi.org/10.1039/c2lc40074j
H. Kimura, T. Yamamoto, H. Sakai, Y. Sakai, T. Fujii, An integrated microfluidic system for long-term perfusion culture and on-line monitoring of intestinal tissue models. Lab Chip 8(5), 741–746 (2008). https://doi.org/10.1039/b717091b
M. Komeya, K. Hayashi, H. Nakamura, H. Yamanaka, H. Sanjo, K. Kojima, T. Ogawa, Pumpless microfluidic system driven by hydrostatic pressure induces and maintains mouse spermatogenesis in vitro. Sci. Rep. 7(1), 15459 (2017). https://doi.org/10.1038/s41598-017-15799-3
S. Lee, J. Lim, J. Yu, J. Ahn, Y. Lee, N.L. Jeon, Engineering tumor vasculature on an injection-molded plastic array 3D culture (IMPACT) platform. Lab Chip 19(12), 2071–2080 (2019). https://doi.org/10.1039/c9lc00148d
M. Marimuthu, S. Kim, Pumpless steady-flow microfluidic chip for cell culture. Anal Biochem. 437(2), 161–163 (2013). https://doi.org/10.1016/j.ab.2013.02.007
T. Mathur, K.A. Singh, NK, R. P., Tsai, S. H., Hein, T. W., Gaharwar, A. K., Jain, A., Organ-on-chips made of blood: endothelial progenitor cells from blood reconstitute vascular thromboinflammation in vessel-chips. Lab Chip 19(15), 2500–2511 (2019). https://doi.org/10.1039/c9lc00469f
I. Meyvantsson, J.W. Warrick, S. Hayes, A. Skoien, D.J. Beebe, Automated cell culture in high density tubeless microfluidic device arrays. Lab Chip 8(5), 717–724 (2008). https://doi.org/10.1039/b715375a
A. Mukherjee, J. Hooks, Z. Nepiyushchikh, J.B. Dixon, Entrainment of Lymphatic Contraction to Oscillatory Flow. Sci. Rep. 9(1), 5840 (2019). https://doi.org/10.1038/s41598-019-42142-9
D.H. Nguyen, S.C. Stapleton, M.T. Yang, S.S. Cha, C.K. Choi, P.A. Galie, C.S. Chen, Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro. Proc. Natl. Acad. Sci. U S A 110(17), 6712–6717 (2013). https://doi.org/10.1073/pnas.1221526110
L.J.Y. Ong, T. Ching, L.H. Chong, S. Arora, H. Li, M. Hashimoto, Y.C. Toh, Self-aligning Tetris-Like (TILE) modular microfluidic platform for mimicking multi-organ interactions. Lab Chip 19(13), 2178–2191 (2019). https://doi.org/10.1039/c9lc00160c
C.E. Owens, A.J. Hart, High-precision modular microfluidics by micromilling of interlocking injection-molded blocks. Lab Chip 18(6), 890–901 (2018). https://doi.org/10.1039/c7lc00951h
D. Park, J. Lee, J.J. Chung, Y. Jung, S.H. Kim, Integrating Organs-on-Chips: Multiplexing, Scaling, Vascularization, and Innervation. Trends Biotechnol. 38(1), 99–112 (2020). https://doi.org/10.1016/j.tibtech.2019.06.006
W.J. Polacheck, M.L. Kutys, J.B. Tefft, C.S. Chen, Microfabricated blood vessels for modeling the vascular transport barrier. Nat. Protoc. 14(5), 1425–1454 (2019). https://doi.org/10.1038/s41596-019-0144-8
C. Probst, S. Schneider, P. Loskill, High-throughput organ-on-a-chip systems: Current status and remaining challenges. Curr. Opin. Biomed. Eng. 6, 33–41 (2018). https://doi.org/10.1016/j.cobme.2018.02.004
L. Richardson, S. Jeong, S. Kim, A. Han, R. Menon, Amnion membrane organ-on-chip: an innovative approach to study cellular interactions. FASEB J 33(8), 8945–8960 (2019). https://doi.org/10.1096/fj.201900020RR
K.S. Ryu, K. Shaikh, E. Goluch, Z. Fan, C. Liu, Micro magnetic stir-bar mixer integrated with parylene microfluidic channels. Lab Chip 4(6), 608–613 (2004). https://doi.org/10.1039/b403305a
E.K. Sackmann, A.L. Fulton, D.J. Beebe, The present and future role of microfluidics in biomedical research. Nature 507(7491), 181–189 (2014). https://doi.org/10.1038/nature13118
A. Sinha, P. Gopinathan, Y.D. Chung, H.Y. Lin, K.H. Li, H.P. Ma, G.B. Lee, An integrated microfluidic platform to perform uninterrupted SELEX cycles to screen affinity reagents specific to cardiovascular biomarkers. Biosens. Bioelectron. 122, 104–112 (2018). https://doi.org/10.1016/j.bios.2018.09.040
W. Song, Q. Wei, W. Liu, T. Liu, J. Yi, N. Sheibani, H.F. Zhang, A combined method to quantify the retinal metabolic rate of oxygen using photoacoustic ophthalmoscopy and optical coherence tomography. Sci. Rep. 4, 6525 (2014). https://doi.org/10.1038/srep06525
S.G.M. Uzel, R.J. Platt, V. Subramanian, T.M. Pearl, C.J. Rowlands, V. Chan, R.D. Kamm, Microfluidic device for the formation of optically excitable, three-dimensional, compartmentalized motor units. Sci. Adv. 2, e1501429 (2016). https://doi.org/10.1126/sciadv.1501429
M.H. Wu, S.B. Huang, Z. Cui, Z. Cui, G.B. Lee, A high throughput perfusion-based microbioreactor platform integrated with pneumatic micropumps for three-dimensional cell culture. Biomed. Microdevices 10(2), 309–319 (2008). https://doi.org/10.1007/s10544-007-9138-3
Y. Xing, M. Nourmohammadzadeh, J.E. Elias, M. Chan, Z. Chen, J.J. McGarrigle, Y. Wang, A pumpless microfluidic device driven by surface tension for pancreatic islet analysis. Biomed. Microdevice 18(5), 80 (2016). https://doi.org/10.1007/s10544-016-0109-4
Y. Yang, P. Fathi, G. Holland, D. Pan, N.S. Wang, M.B. Esch, Pumpless microfluidic devices for generating healthy and diseased endothelia. Lab Chip 19(19), 3212–3219 (2019). https://doi.org/10.1039/c9lc00446g
J. Yeste, M. Garcia-Ramirez, X. Illa, A. Guimera, C. Hernandez, R. Simo, R. Villa, A compartmentalized microfluidic chip with crisscross microgrooves and electrophysiological electrodes for modeling the blood-retinal barrier. Lab Chip 18(1), 95–105 (2017). https://doi.org/10.1039/c7lc00795g
H.G. Yi, Y.H. Jeong, Y. Kim, Y.J. Choi, H.E. Moon, S.H. Park, D.W. Cho, A bioprinted human-glioblastoma-on-a-chip for the identification of patient-specific responses to chemoradiotherapy. Nat. Biomed. Eng. 3(7), 509–519 (2019). https://doi.org/10.1038/s41551-019-0363-x
P.K. Yuen, Fluid control in microfluidic devices using a fluid conveyance extension and an absorbent microfluidic flow modulator. Lab Chip 13(9), 1737–1742 (2013). https://doi.org/10.1039/c3lc40956b
B. Zhang, A. Korolj, B.F.L. Lai, M. Radisic, Advances in organ-on-a-chip engineering. Nat. Rev. Mater. 3(8), 257–278 (2018). https://doi.org/10.1038/s41578-018-0034-7
Acknowledgments
This work was supported in part by James Tronolone’s appointment to the Research Participation Program at US Food and Drug Administration (FDA) administered by the Oak Ridge Institute for Science and Education through the US Department of Education and FDA. This work was also partially supported by the research funds from the Division of Cellular and Gene Therapies at FDA.
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Tronolone, J.J., Lam, J., Agrawal, A. et al. Pumpless, modular, microphysiological systems enabling tunable perfusion for long-term cultivation of endothelialized lumens. Biomed Microdevices 23, 25 (2021). https://doi.org/10.1007/s10544-021-00562-3
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DOI: https://doi.org/10.1007/s10544-021-00562-3