Micropatterned coculture of primary human hepatocytes and supportive cells for the study of hepatotropic pathogens

Nature Protocols

Volumn 10, Issue 12, 2015, Pages 2027-2053

  March, Sandra ^a,b   Ramanan, Vyas ^b   Trehan, Kartik ^b   Ng, Shengyong ^b,c   Galstian, Ani ^a,b   Gural, Nil ^b   Scull, Margaret A ^d   Shlomai, Amir ^d   Mota, Maria M ^e   Fleming, Heather E ^b   Khetani, Salman R ^f   Rice, Charles M ^d   Bhatia, Sangeeta N ^a,b,g,h,i  

^a BROAD INSTITUTE OF MIT AND HARVARD   (United States)

^b MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

^c Massachusetts Institute of Technology   (United States)

^d ROCKEFELLER UNIVERSITY   (United States)

^e UNIVERSITY OF LISBON   (Portugal)

^f Department of Mechanical Engineering   (United States)

^g BRIGHAM AND WOMEN S HOSPITAL   (United States)

^h Department of Electrical Engineering and Computer Science   (United States)

ⁱ HOWARD HUGHES MEDICAL INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

COVALENTLY CLOSED CIRCULAR DNA; DNA; ENTECAVIR; HEPATITIS B SURFACE ANTIGEN; HEPATITIS B(E) ANTIGEN; INTERFERON; RED FLUORESCENT PROTEIN; UNCLASSIFIED DRUG; VIRUS RNA;

ANTIVIRAL SUSCEPTIBILITY; ARTICLE; COCULTURE; FIBROBLAST; HEPATITIS B VIRUS; HEPATITIS C VIRUS; HOST PATHOGEN INTERACTION; HUMAN; LIVER CELL CULTURE; MICROPATTERNED COCULTURE; MINICHROMOSOME; NONHUMAN; NUCLEAR LOCALIZATION SIGNAL; PLASMODIUM FALCIPARUM; PLASMODIUM VIVAX; PRIORITY JOURNAL; SCHIZONT; SPOROZOITE; VIRUS CELL INTERACTION; ANIMAL; CELL CULTURE; CELL LINE; CYTOLOGY; HEPACIVIRUS; HEPATITIS B; HEPATITIS C; LIVER CELL; MALARIA; METABOLISM; MOUSE; PARASITOLOGY; PHYSIOLOGY; PLASMODIUM; PROCEDURES; TISSUE MICROARRAY; VIROLOGY;

ANIMALS; CELL LINE; CELLS, CULTURED; COCULTURE TECHNIQUES; FIBROBLASTS; HEPACIVIRUS; HEPATITIS B; HEPATITIS B VIRUS; HEPATITIS C; HEPATOCYTES; HOST-PATHOGEN INTERACTIONS; HUMANS; MALARIA; MICE; PLASMODIUM; PLASMODIUM FALCIPARUM; PLASMODIUM VIVAX; TISSUE ARRAY ANALYSIS;

EID: 84947934013     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2015.128     Document Type: Article

References (97)

1. Lavanchy, D. The global burden of hepatitis C. Liver Int. 29 (suppl. 1), 74-81 (2009).
2. Ganem, D. & Prince, A.M. Hepatitis B virus infection-natural history and clinical consequences. N. Engl. J. Med. 350, 1118-1129 (2004).
3. Razavi, H. et al. Chronic hepatitis C virus (HCV) disease burden and cost in the United States. Hepatology 57, 2164-2170 (2013).
4. Alonso, P.L. et al. A research agenda for malaria eradication: drugs. PLoS Med. 8, e1000402 (2011b).
5. Wells, T.N., Alonso, P.L. & Gutteridge, W.E. New medicines to improve control and contribute to the eradication of malaria. Nat. Rev. Drug Discov. 8, 879-891 (2009).
6. Wells, T.N., Burrows, J.N. & Baird, J.K. Targeting the hypnozoite reservoir of Plasmodium vivax: the hidden obstacle to malaria elimination. Trends Parasitol. 26, 145-151 (2010).
7. Billerbeck, E., de Jong, Y., Dorner, M., de la Fuente, C. & Ploss, A. Animal models for hepatitis C. Curr. Top. Microbiol. Immunol. 369, 49-86 (2013).
8. Kaushansky, A., Mikolajczak, S.A., Vignali, M. & Kappe, S.H. Of men in mice: the success and promise of humanized mouse models for human malaria parasite infections. Cell Microbiol. 16, 602-611 (2014).
9. Lohmann, V. & Bartenschlager, R. On the history of hepatitis C virus cell culture systems. J. Med. Chem. 57, 1627-1642 (2014).
10. Farkas, D. & Tannenbaum, S.R. In vitro methods to study chemically-induced hepatotoxicity: a literature review. Curr. Drug Metab. 6, 111-125 (2005).
11. Godoy, P. et al. Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch. Toxicol. 87, 1315-1530 (2013).
12. Guillouzo, A. Liver cell models in in vitro toxicology. Environ. Health Perspect. 106 (suppl. 2), 511-532 (1998).
13. Khetani, S.R. et al. Microengineered liver tissues for drug testing. J. Lab. Autom. 20, 216-250 (2015).
14. Guillouzo, A. & Guguen-Guillouzo, C. Evolving concepts in liver tissue modeling and implications for in vitro toxicology. Expert Opin. Drug Metab. Toxicol. 4, 1279-1294 (2008).
15. LeCluyse, E.L. Human hepatocyte culture systems for the in vitro evaluation of cytochrome P450 expression and regulation. Eur. J. Pharm. Sci. 13, 343-368 (2001).
16. Lagaye, S. et al. Effcient replication of primary or culture hepatitis C virus isolates in human liver slices: a relevant ex vivo model of liver infection. Hepatology 56, 861-872 (2012).
17. van Midwoud, P.M., Merema, M.T., Verweij, N., Groothuis, G.M. & Verpoorte, E. Hydrogel embedding of precision-cut liver slices in a microfuidic device improves drug metabolic activity. Biotechnol. Bioeng. 108, 1404-1412 (2011).
18. Gerets, H.H. et al. Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol. Toxicol. 28, 69-87 (2012).
19. Wilkening, S., Stahl, F. & Bader, A. Comparison of primary human hepatocytes and hepatoma cell line Hepg2 with regard to their biotransformation properties. Drug Metab. Dispos. 31, 1035-1042 (2003).
20. Andersson, T.B., Kanebratt, K.P. & Kenna, J.G. The HepaRG cell line: a unique in vitro tool for understanding drug metabolism and toxicology in human. Expert Opin. Drug Metab. Toxicol. 8, 909-920 (2012).
21. Le Vee, M. et al. Functional expression of sinusoidal and canalicular hepatic drug transporters in the differentiated human hepatoma HepaRG cell line. Eur. J. Pharm. Sci. 28, 109-117 (2006).
22. Le Vee, M., Noel, G., Jouan, E., Stieger, B. & Fardel, O. Polarized expression of drug transporters in differentiated human hepatoma HepaRG cells. Toxicol. In Vitro 27, 1979-1986 (2013).
23. Lubberstedt, M. et al. HepaRG human hepatic cell line utility as a surrogate for primary human hepatocytes in drug metabolism assessment in vitro. J. Pharmacol. Toxicol. Methods 63, 59-68 (2011).
24. Szabo, M., Veres, Z., Baranyai, Z., Jakab, F. & Jemnitz, K. Comparison of human hepatoma HepaRG cells with human and rat hepatocytes in uptake transport assays in order to predict a risk of drug-induced hepatotoxicity. PLoS ONE 8, e59432 (2013).
25. Xu, J.J. et al. Cellular imaging predictions of clinical drug-induced liver injury. Toxicol. Sci. 105, 97-105 (2008).
26. Bi, Y.A., Kazolias, D. & Duignan, D.B. Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport. Drug Metab. Dispos. 34, 1658-1665 (2006).
27. Dunn, J.C., Yarmush, M.L., Koebe, H.G. & Tompkins, R.G. Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich confguration. FASEB J. 3, 174-177 (1989).
28. Khetani, S.R. & Bhatia, S.N. Microscale culture of human liver cells for drug development. Nat. Biotechnol. 26, 120-126 (2008).
29. Sivaraman, A. et al. A microscale in vitro physiological model of the liver: predictive screens for drug metabolism and enzyme induction. Curr. Drug Metab. 6, 569-591 (2005).
30. LeCluyse, E.L., Witek, R.P., Andersen, M.E. & Powers, M.J. Organotypic liver culture models: meeting current challenges in toxicity testing. Crit. Rev. Toxicol. 42, 501-548 (2012).
31. Ramsden, D., Zhou, J. & Tweedie, D.J. Determination of a degradation constant for CYP3A4 by direct suppression of mRNA in a novel human hepatocyte model, HepatoPac. Drug Metab. Dispos. 43, 1307-1315 (2015).
32. Si-Tayeb, K. et al. Highly effcient generation of human hepatocyte-like cells from induced pluripotent stem cells. Hepatology 51, 297-305 (2010).
33. Schwartz, R.E., Fleming, H.E., Khetani, S.R. & Bhatia, S.N. Pluripotent stem cell-derived hepatocyte-like cells. Biotechnol. Adv. 32, 504-513 (2014).
34. Cai, J. et al. in StemBook 1.52.1 (The Stem Cell Research Community) (Harvard Stem Cell Institute, 2012).
35. Berger, D.R., Ware, B.R., Davidson, M.D., Allsup, S.R. & Khetani, S.R. Enhancing the functional maturity of induced pluripotent stem cell-derived human hepatocytes by controlled presentation of cell-cell interactions in vitro. Hepatology 61, 1370-1381 (2015).
36. Shan, J. et al. Identifcation of small molecules for human hepatocyte expansion and iPS differentiation. Nat. Chem. Biol. 9, 514-520 (2013).
37. Bhatia, S.N. & Ingber, D.E. Microfuidic organs-on-chips. Nat. Biotechnol. 32, 760-772 (2014).
38. Gripon, P. et al. Infection of a human hepatoma cell line by hepatitis B virus. Proc. Natl. Acad. Sci. USA 99, 15655-15660 (2002).
39. Rehermann, B. & Nascimbeni, M. Immunology of hepatitis B virus and hepatitis C virus infection. Nat. Rev. Immunol. 5, 215-229 (2005).
40. Gripon, P. et al. Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J. Virol. 62, 4136-4143 (1988).
41. Banaudha, K. et al. Primary hepatocyte culture supports hepatitis C virus replication: a model for infection-associated hepatocarcinogenesis. Hepatology 51, 1922-1932 (2010).
42. Buck, M. Direct infection and replication of naturally occurring hepatitis C virus genotypes 1, 2, 3 and 4 in normal human hepatocyte cultures. PLoS ONE 3, e2660 (2008).
43. Harding, M.J. et al. An implantable vascularized protein gel construct that supports human fetal hepatoblast survival and infection by hepatitis C virus in mice. PLoS ONE 5, e9987 (2010).
44. Chattopadhyay, R. et al. Establishment of an in vitro assay for assessing the effects of drugs on the liver stages of Plasmodium vivax malaria. PLoS ONE 5, e14275 (2010).
45. + T cell immunity. Science 334, 475 (2011).
46. Hollingdale, M.R., Collins, W.E. & Campbell, C.C. In vitro culture of exoerythrocytic parasites of the North Korean strain of Plasmodium vivax in hepatoma cells. Am. J. Trop. Med. Hyg. 35, 275-276 (1986).
47. Karnasuta, C. et al. Complete development of the liver stage of Plasmodium falciparum in a human hepatoma cell line. Am. J. Trop. Med. Hyg. 53, 607-611 (1995).
48. Sattabongkot, J. et al. Establishment of a human hepatocyte line that supports in vitro development of the exo-erythrocytic stages of the malaria parasites Plasmodium falciparum and P. vivax. Am. J. Trop. Med. Hyg. 74, 708-715 (2006).
49. Yokoo, H. et al. Proteomic signature corresponding to α-fetoprotein expression in liver cancer cells. Hepatology 40, 609-617 (2004).
50. Dembele, L. et al. Persistence and activation of malaria hypnozoites in long-term primary hepatocyte cultures. Nat. Med. 20, 307-312 (2014).
51. Mazier, D. et al. Complete development of hepatic stages of Plasmodium falciparum in vitro. Science 227, 440-442 (1985).
52. Mazier, D. et al. Cultivation of the liver forms of Plasmodium vivax in human hepatocytes. Nature 307, 367-369 (1984).
53. van Schaijk, B.C. et al. Gene disruption of Plasmodium falciparum p52 results in attenuation of malaria liver stage development in cultured primary human hepatocytes. PLoS ONE 3, e3549 (2008).
54. Yalaoui, S. et al. Scavenger receptor BI boosts hepatocyte permissiveness to Plasmodium infection. Cell Host Microbe 4, 283-292 (2008).
55. Bhatia, S.N., Balis, U.J., Yarmush, M.L. & Toner, M. Effect of cell-cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells. FASEB J. 13, 1883-1900 (1999).
56. March, S. et al. A microscale human liver platform that supports the hepatic stages of Plasmodium falciparum and vivax. Cell Host Microbe 14, 104-115 (2013).
57. Ploss, A. et al. Persistent hepatitis C virus infection in microscale primary human hepatocyte cultures. Proc. Natl. Acad. Sci. USA 107, 3141-3145 (2010).
58. Shlomai, A. et al. Modeling host interactions with hepatitis B virus using primary and induced pluripotent stem cell-derived hepatocellular systems. Proc. Natl. Acad. Sci. USA 111, 12193-12198 (2014).
59. Guguen-Guillouzo, C. et al. Maintenance and reversibility of active albumin secretion by adult rat hepatocytes co-cultured with another liver epithelial cell type. Exp. Cell Res. 143, 47-54 (1983).
60. Rheinwald, J.G. & Green, H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6, 331-343 (1975).
61. Khetani, S.R., Szulgit, G., Del Rio, J.A., Barlow, C. & Bhatia, S.N. Exploring interactions between rat hepatocytes and nonparenchymal cells using gene expression profling. Hepatology 40, 545-554 (2004).
62. Leclercq, L. et al. Which human metabolites have we MIST? Retrospective analysis, practical aspects, and perspectives for metabolite identifcation and quantifcation in pharmaceutical development. Chem. Res. Toxicol. 22, 280-293 (2009).
63. Olson, H. et al. Concordance of the toxicity of pharmaceuticals in humans and in animals. Regul. Toxicol. Pharmacol. 32, 56-67 (2000).
64. Shih, H., Pickwell, G.V., Guenette, D.K., Bilir, B. & Quattrochi, L.C. Species differences in hepatocyte induction of CYP1A1 and CYP1A2 by omeprazole. Hum. Exp. Toxicol. 18, 95-105 (1999).
65. Wang, W.W., Khetani, S.R., Krzyzewski, S., Duignan, D.B. & Obach, R.S. Assessment of a micropatterned hepatocyte coculture system to generate major human excretory and circulating drug metabolites. Drug Metab. Dispos. 38, 1900-1905 (2010).
66. Khetani, S.R. et al. Use of micropatterned cocultures to detect compounds that cause drug-induced liver injury in humans. Toxicol. Sci. 132, 107-117 (2013).
67. Chan, T.S., Yu, H., Moore, A., Khetani, S.R. & Tweedie, D. Meeting the challenge of predicting hepatic clearance of compounds slowly metabolized by cytochrome P450 using a novel hepatocyte model, HepatoPac. Drug Metab. Dispos. 41, 2024-2032 (2013).
68. Trask, O.J. Jr., Moore, A. & LeCluyse, E.L. A micropatterned hepatocyte coculture model for assessment of liver toxicity using high-content imaging analysis. Assay Drug Dev. Technol. 12, 16-27 (2014).
69. Ramsden, D., Tweedie, D.J., Chan, T.S., Taub, M.E. & Li, Y. Bridging in vitro and in vivo metabolism and transport of faldaprevir in human using a novel cocultured human hepatocyte system, HepatoPac. Drug Metab. Dispos. 42, 394-406 (2014).
70. Ng, S. et al. Hypoxia promotes liver-stage malaria infection in primary human hepatocytes in vitro. Dis. Model Mech. 7, 215-224 (2014).
71. Ukairo, O. et al. Long-term stability of primary rat hepatocytes in micropatterned cocultures. J. Biochem. Mol. Toxicol. 27, 204-212 (2013).
72. Aoyama, S., Lambirth, S. & Khetani, S.R. A long-term culture model for primary hepatoyctes from Cynomolgus hepatocytes. Drug Metab. Rev. 43, 124 (2011).
73. Dembele, L. et al. Towards an in vitro model of Plasmodium hypnozoites suitable for drug discovery. PLoS ONE 6, e18162 (2011).
74. Davidson, M.D., Lehrer, M. & Khetani, S.R. Hormone and drug-mediated modulation of glucose metabolism in a microscale model of the human liver. Tissue Eng. Part C Methods 21, 716-725 (2015).
75. Qin, D., Xia, Y. & Whitesides, G.M. Soft lithography for micro-and nanoscale patterning. Nat. Protoc. 5, 491-502 (2010).
76. March, S., Hui, E.E., Underhill, G.H., Khetani, S. & Bhatia, S.N. Microenvironmental regulation of the sinusoidal endothelial cell phenotype in vitro. Hepatology 50, 920-928 (2009).
77. Chen, A.A., Khetani, S.R., Lee, S., Bhatia, S.N. & Van Vliet, K.J. Modulation of hepatocyte phenotype in vitro via chemomechanical tuning of polyelectrolyte multilayers. Biomaterials 30, 1113-1120 (2009).
78. Khetani, S.R., Chen, A.A., Ranscht, B. & Bhatia, S.N. T-cadherin modulates hepatocyte functions in vitro. FASEB J. 22, 3768-3775 (2008).
79. Meis, J.F. et al. Fine structure of the malaria parasite Plasmodium falciparum in human hepatocytes in vitro. Cell Tissue Res. 244, 345-350 (1986).
80. Shortt, H. History of recent researches on tissue phases of the malaria parasite at the London School of Hygiene and Tropical Medicine. Trans. R. Soc. Trop. Med. Hyg. 45, 175-188 (1951).
81. Shortt, H.E., Garnham, P.C. & Malamos, B. The pre-erythrocytic stage of mammalian malaria. Br. Med. J. 1, 192-194 (1948).
82. Vaughan, A.M. et al. Complete Plasmodium falciparum liver-stage development in liver-chimeric mice. J. Clin. Invest. 122, 3618-3628 (2012).
83. Todaro, G.J. & Green, H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J. Cell Biol. 17, 299-313 (1963).
84. Bhatia, S.N., Yarmush, M.L. & Toner, M. Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fbroblasts. J. Biomed. Mater. Res. 34, 189-199 (1997).
85. Zhong, J. et al. Robust hepatitis C virus infection in vitro. Proc. Natl. Acad. Sci. USA 102, 9294-9299 (2005).
86. Sells, M.A., Chen, M.L. & Acs, G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc. Natl. Acad. Sci. USA 84, 1005-1009 (1987).
87. Lindenbach, B.D. et al. Complete replication of hepatitis C virus in cell culture. Science 309, 623-626 (2005).
88. Jones, C.T. et al. Real-time imaging of hepatitis C virus infection using a fuorescent cell-based reporter system. Nat. Biotechnol. 28, 167-171 (2010).
89. Albuquerque, S.S. et al. Host cell transcriptional profling during malaria liver stage infection reveals a coordinated and sequential set of biological events. BMC Genomics 10, 270 (2009).
90. Hoffman, S.L. et al. Development of a metabolically active, non-replicating sporozoite vaccine to prevent Plasmodium falciparum malaria. Hum. Vaccin. 6, 97-106 (2010).
91. Renia, L. et al. Malaria sporozoite penetration. A new approach by double staining. J. Immunol. Methods 112, 201-205 (1988).
92. Silvie, O. et al. Hepatocyte CD81 is required for Plasmodium falciparum and Plasmodium yoelii sporozoite infectivity. Nat. Med. 9, 93-96 (2003).
93. Marukian, S. et al. Cell culture-produced hepatitis C virus does not infect peripheral blood mononuclear cells. Hepatology 48, 1843-1850 (2008).
94. Glebe, D. et al. Pre-s1 antigen-dependent infection of Tupaia hepatocyte cultures with human hepatitis B virus. J. Virol. 77, 9511-9521 (2003).
95. Portugal, S., Drakesmith, H. & Mota, M.M. Superinfection in malaria: Plasmodium shows its iron will. EMBO Rep. 12, 1233-1242 (2011).
96. Arzberger, S., Hosel, M. & Protzer, U. Apoptosis of hepatitis B virus-infected hepatocytes prevents release of infectious virus. J. Virol. 84, 11994-12001 (2010).
97. Reed, L.J. & Muench, H. A simple method of estimating ffty percent endpoints. Am. J. Hyg. 27, 493-497 (1938).