Abstract
Helicobacter research classically uses fixed human tissue, animal models or cancer cell lines. Each of these study objects has its advantages and has brought central insights into the infection process. Nevertheless, in model systems for basic and medical research, there is a gap between two-dimensional and most often transformed cell cultures and three-dimensional, highly organized tissues. In recent years, stem cell research has provided the means to fill this gap. The identification of the niche factors that support growth, expansion and differentiation of stem cells in vitro has allowed the development of three-dimensional culture systems called organoids. Gastric organoids are grown from gastric stem cells and are organized epithelial structures that comprise all the differentiated cell types of the stomach. They can be expanded without apparent limitation and are amenable to a wide range of standard laboratory techniques. Here, we review different stem cell-derived organoid model systems useful for Helicobacter pylori research and outline their advantages for infection studies.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Aebischer T, Bumann D, Epple HJ et al (2008) Correlation of T cell response and bacterial clearance in human volunteers challenged with Helicobacter pylori revealed by randomised controlled vaccination with Ty21a-based Salmonella vaccines. Gut 57:1065–1072. doi:10.1136/gut.2007.145839
Andersson-Rolf A, Fink J, Mustata RC, Koo B-K (2014) A video protocol of retroviral infection in primary intestinal organoid culture. J Vis Exp. doi:10.3791/51765
Arnold K, Sarkar A, Yram MA et al (2011) Sox2+ adult stem and progenitor cells are important for tissue regeneration and survival of mice. Cell Stem Cell 9:317–329. doi:10.1016/j.stem.2011.09.001
Backert S, Naumann M (2010) What a disorder: proinflammatory signaling pathways induced by Helicobacter pylori. Trends Microbiol 18(11):479–486. doi:10.1016/j.tim.2010.08.003
Backert S, Ziska E, Brinkmann V et al (2000) Translocation of the Helicobacter pylori CagA protein in gastric epithelial cells by a type IV secretion apparatus. Cell Microbiol 2:155–164. doi:10.1046/j.1462-5822.2000.00043.x
Backert S, Tegtmeyer N, Fischer W (2015) Composition, structure and function of the Helicobacter pylori cag pathogenicity island encoded type IV secretion system. Future Microbiol 10(6):955–965. doi:10.2217/fmb.15.32
Barker N, Huch M, Kujala P et al (2010) Lgr5+ ve stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell 6:25–36. doi:10.1016/j.stem.2009.11.013
Bartfeld S, Clevers H (2015) Organoids as model for infectious diseases: culture of human and murine stomach organoids and microinjection of Helicobacter pylori. J Vis Exp. doi:10.3791/53359
Bartfeld S, Hess S, Bauer B et al (2010) High-throughput and single-cell imaging of NF-kappaB oscillations using monoclonal cell lines. BMC Cell Biol 11:21. doi:10.1186/1471-2121-11-21
Bartfeld S, Bayram T, van de Wetering M et al (2015) In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. Gastroenterology 148(126–136):e6. doi:10.1053/j.gastro.2014.09.042
Bartfeld S, Koo BK (2016). Adult gastric stem cells and their niches. WIREs Dev Biol. doi:10.1002/wdev.261
Bertaux-Skeirik N, Feng R, Schumacher MA et al (2015) CD44 plays a functional role in Helicobacter pylori-induced epithelial cell proliferation. PLoS Pathog 11:e1004663. doi:10.1371/journal.ppat.1004663
Blaser MJ, Kirschner D (1999) Dynamics of Helicobacter pylori colonization in relation to the host response. Proc Natl Acad Sci 96:8359–8364. doi:10.1073/pnas.96.15.8359
Blaser MJ, Perez-Perez GI, Kleanthous H et al (1995) Infection with Helicobacter pylori strains possessing cagA is associated with an increased risk of developing adenocarcinoma of the stomach. Cancer Res 55:2111–2115
Boj SF, Hwang C-I, Baker LA et al (2015) Organoid models of human and mouse ductal pancreatic cancer. Cell 160:324–338. doi:10.1016/j.cell.2014.12.021
Boxberger H-J, Sessler MJ, Grausam MC et al (1997) Isolation and culturing of highly polarized primary epithelial cells from normal human stomach (antrum) as spheroid-like vesicles. Methods Cell Sci 19:169–178. doi:10.1023/A:1009751913391
Bredemeyer AJ, Geahlen JH, Weis VG et al (2009) The gastric epithelial progenitor cell niche and differentiation of the zymogenic (chief) cell lineage. Dev Biol 325:211–224. doi:10.1016/j.ydbio.2008.10.025
Choi E, Roland JT, Barlow BJ et al (2014) Cell lineage distribution atlas of the human stomach reveals heterogeneous gland populations in the gastric antrum. Gut 63:1711–1720. doi:10.1136/gutjnl-2013-305964
Churin Y, Al-Ghoul L, Kepp O et al (2003) Helicobacter pylori CagA protein targets the c-Met receptor and enhances the motogenic response. J Cell Biol 161:249–255. doi:10.1083/jcb.200208039
Covacci A, Censini S, Bugnoli M et al (1993) Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci 90:5791–5795
Dedhia PH, Bertaux-Skeirik N, Zavros Y, Spence JR (2016) Organoid models of human gastrointestinal development and disease. Gastroenterology 150:1098–1112. doi:10.1053/j.gastro.2015.12.042
Demitrack ES, Gifford GB, Keeley TM et al (2015) Notch signaling regulates gastric antral LGR5 stem cell function. EMBO J 34:2522–2536. doi:10.15252/embj.201490583
Drost J, van Jaarsveld RH, Ponsioen B et al (2015) Sequential cancer mutations in cultured human intestinal stem cells. Nature 521:43–47. doi:10.1038/nature14415
Fatehullah A, Tan SH, Barker N (2016) Organoids as an in vitro model of human development and disease. Nat Cell Biol 18:246–254. doi:10.1038/ncb3312
Feng R, Aihara E, Kenny S et al (2014) Indian hedgehog mediates gastrin-induced proliferation in stomach of adult mice. Gastroenterology 147(655–666):e9. doi:10.1053/j.gastro.2014.05.006
Fujii M, Shimokawa M, Date S et al (2016) A colorectal tumor organoid library demonstrates progressive loss of niche factor requirements during tumorigenesis. Cell Stem Cell. doi:10.1016/j.stem.2016.04.003
Goodlad RA, Wilson TJ, Lenton W et al (1987) Proliferative effects of urogastrone-EGF on the intestinal epithelium. Gut 28:37–43
Hayakawa Y, Ariyama H, Stancikova J et al (2015a) Mist1 expressing gastric stem cells maintain the normal and neoplastic gastric epithelium and are supported by a perivascular stem cell niche. Cancer Cell 28:800–814. doi:10.1016/j.ccell.2015.10.003
Hayakawa Y, Jin G, Wang H et al (2015b) CCK2R identifies and regulates gastric antral stem cell states and carcinogenesis. Gut 64:544–553. doi:10.1136/gutjnl-2014-307190
Hessey SJ, Spencer J, Wyatt JI et al (1990) Bacterial adhesion and disease activity in Helicobacter associated chronic gastritis. Gut 31:134–138
Huang JY, Sweeney EG, Sigal M et al (2015) Chemodetection and destruction of host urea allows Helicobacter pylori to locate the epithelium. Cell Host Microbe 18:147–156. doi:10.1016/j.chom.2015.07.002
Huch M, Koo B-K (2015) Modeling mouse and human development using organoid cultures. Dev Camb Engl 142:3113–3125. doi:10.1242/dev.118570
Huch M, Bonfanti P, Boj SF et al (2013) Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis. EMBO J 32:2708–2721. doi:10.1038/emboj.2013.204
Karam SM (1993) Dynamics of epithelial cells in the corpus of the mouse stomach. IV. Bidirectional migration of parietal cells ending in their gradual degeneration and loss. Anat Rec 236:314–332. doi:10.1002/ar.1092360205
Karam SM, Leblond CP (1993a) Dynamics of epithelial cells in the corpus of the mouse stomach. III. Inward migration of neck cells followed by progressive transformation into zymogenic cells. Anat Rec 236:297–313. doi:10.1002/ar.1092360204
Karam SM, Leblond CP (1993b) Dynamics of epithelial cells in the corpus of the mouse stomach. II. Outward migration of pit cells. Anat Rec 236:280–296. doi:10.1002/ar.1092360203
Karthaus WR, Iaquinta PJ, Drost J et al (2014) Identification of multipotent luminal progenitor cells in human prostate organoid cultures. Cell 159:163–175. doi:10.1016/j.cell.2014.08.017
Katano T, Ootani A, Mizoshita T et al (2013) Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche. Biochem Biophys Res Commun 432:558–563. doi:10.1016/j.bbrc.2013.02.051
Kessler M, Hoffmann K, Brinkmann V et al (2015) The Notch and Wnt pathways regulate stemness and differentiation in human fallopian tube organoids. Nat Commun 6:8989. doi:10.1038/ncomms9989
Koo B-K, Stange DE, Sato T et al (2012) Controlled gene expression in primary Lgr5 organoid cultures. Nat Methods 9:81–83. doi:10.1038/nmeth.1802
Kretzschmar K, Watt FM (2012) Lineage tracing. Cell 148:33–45. doi:10.1016/j.cell.2012.01.002
Lancaster MA, Knoblich JA (2014) Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345:1247125. doi:10.1126/science.1247125
Lee A (1998) Animal models for host-pathogen interaction studies. Br Med Bull 54:163–173
Lee ER, Leblond CP (1985a) Dynamic histology of the antral epithelium in the mouse stomach: IV. Ultrastructure and renewal of gland cells. Am J Anat 172:241–259
Lee ER, Leblond CP (1985b) Dynamic histology of the antral epithelium in the mouse stomach: II. Ultrastructure and renewal of isthmal cells. Am J Anat 172:205–224. doi:10.1002/aja.1001720304
Lee ER, Trasler J, Dwivedi S, Leblond CP (1982) Division of the mouse gastric mucosa into zymogenic and mucous regions on the basis of gland features. Am J Anat 164:187–207. doi:10.1002/aja.1001640302
Li VSW, Ng SS, Boersema PJ et al (2012) Wnt signaling through inhibition of β-Catenin degradation in an intact Axin1 complex. Cell 149:1245–1256. doi:10.1016/j.cell.2012.05.002
Li X, Nadauld L, Ootani A et al (2014) Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture. Nat Med 20:769–777. doi:10.1038/nm.3585
Marshall BJ, Armstrong JA, McGechie DB, Glancy RJ (1985) Attempt to fulfil Koch’s postulates for pyloric Campylobacter. Med J Aust 142:436–439
Matano M, Date S, Shimokawa M et al (2015) Modeling colorectal cancer using CRISPR-Cas9–mediated engineering of human intestinal organoids. Nat Med. doi:10.1038/nm.3802
McCracken KW, Howell JC, Wells JM, Spence JR (2011) Generating human intestinal tissue from pluripotent stem cells in vitro. Nat Protoc 6:1920–1928. doi:10.1038/nprot.2011.410
McCracken KW, Catá EM, Crawford CM et al (2014) Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature 516:400–404. doi:10.1038/nature13863
Mills JC, Sansom OJ (2015) Reserve stem cells: differentiated cells reprogram to fuel repair, metaplasia, and neoplasia in the adult gastrointestinal tract. Sci Signal 8:re8. doi:10.1126/scisignal.aaa7540
Mueller D, Tegtmeyer N, Brandt S, Yamaoka Y, De Poire E, Sgouras D, Wessler S, Torres J, Smolka A, Backert S (2012) c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains. J Clin Invest 122:1553–1566. doi:10.1172/JCI61143
Noguchi TK, Ninomiya N, Sekine M et al (2015) Generation of stomach tissue from mouse embryonic stem cells. Nat Cell Biol 17:984–993. doi:10.1038/ncb3200
Noto JM, Romero-Gallo J, Piazuelo MB, Peek RM (2016) The Mongolian Gerbil: a robust model of Helicobacter pylori-induced gastric inflammation and cancer. Methods Mol Biol Clifton NJ 1422:263–280. doi:10.1007/978-1-4939-3603-8_24
Nyeng P, Norgaard GA, Kobberup S, Jensen J (2007) FGF10 signaling controls stomach morphogenesis. Dev Biol 303:295–310. doi:10.1016/j.ydbio.2006.11.017
Ootani A, Toda S, Fujimoto K, Sugihara H (2000) An air-liquid interface promotes the differentiation of gastric surface mucous cells (GSM06) in culture. Biochem Biophys Res Commun 271:741–746. doi:10.1006/bbrc.2000.2673
Ootani A, Toda S, Fujimoto K, Sugihara H (2003) Foveolar differentiation of mouse gastric mucosa in vitro. Am J Pathol 162:1905–1912. doi:10.1016/S0002-9440(10)64324-6
Ootani A, Li X, Sangiorgi E et al (2009) Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche. Nat Med 15:701–706. doi:10.1038/nm.1951
Posselt G, Backert S, Wessler S (2013) The functional interplay of Helicobacter pylori factors with gastric epithelial cells induces a multi-step process in pathogenesis. Cell Commun Signal CCS 11:77. doi:10.1186/1478-811x-11-77
Qiao XT, Ziel JW, McKimpson W et al (2007) Prospective identification of a multilineage progenitor in murine stomach epithelium. Gastroenterology 133(1989–1998):e3. doi:10.1053/j.gastro.2007.09.031
Radulescu S, Ridgway RA, Cordero J et al (2013) Acute WNT signalling activation perturbs differentiation within the adult stomach and rapidly leads to tumour formation. Oncogene 32:2048–2057. doi:10.1038/onc.2012.224
Richter-Dahlfors A, Heczko U, Meloche RM et al (1998) Helicobacter pylori-infected human antral primary cell cultures: effect on gastrin cell function. Am J Physiol 275:G393–G401
Sato T, Clevers H (2013) Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science 340:1190–1194. doi:10.1126/science.1234852
Sato T, Vries RG, Snippert HJ et al (2009) Single Lgr5 stem cells build crypt villus structures in vitro without a mesenchymal niche. Nature 459:262–265. doi:10.1038/nature07935
Sato T, Stange DE, Ferrante M et al (2011) Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and barrett’s epithelium. Gastroenterology 141:1762–1772. doi:10.1053/j.gastro.2011.07.050
Scanu T, Spaapen RM, Bakker JM et al (2015) Salmonella manipulation of host signaling pathways provokes cellular transformation associated with gallbladder carcinoma. Cell Host Microbe 17:763–774. doi:10.1016/j.chom.2015.05.002
Schlaermann P, Toelle B, Berger H et al (2016) A novel human gastric primary cell culture system for modelling Helicobacter pylori infection in vitro. Gut. doi:10.1136/gutjnl-2014-307949
Schreiber S, Konradt M, Groll C et al (2004) The spatial orientation of Helicobacter pylori in the gastric mucus. Proc Natl Acad Sci USA 101:5024–5029. doi:10.1073/pnas.0308386101
Schumacher MA, Aihara E, Feng R, et al (2015a) The use of murine-derived fundic organoids in studies of gastric physiology: the use of fundic organoids. J Physiol. doi:10.1113/jphysiol.2014.283028
Schumacher MA, Feng R, Aihara E et al (2015b) Helicobacter pylori-induced sonic hedgehog expression is regulated by NFκB pathway activation: the use of a novel in vitro model to study epithelial response to infection. Helicobacter 20:19–28. doi:10.1111/hel.12152
Schwank G, Andersson-Rolf A, Koo B-K et al (2013a) Generation of BAC transgenic epithelial organoids. PLoS ONE 8:e76871. doi:10.1371/journal.pone.0076871
Schwank G, Koo B-K, Sasselli V et al (2013b) Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. Cell Stem Cell 13:653–658. doi:10.1016/j.stem.2013.11.002
Selbach M, Moese S, Hauck CR, Meyer TF, Backert S (2002) Src is the kinase of the Helicobacter pylori CagA protein in vitro and in vivo. J Biol Chem 277:6775–6778. doi:10.1074/jbc.C100754200
Shimoyama T, Crabtree J (1998) Bacterial factors and immune pathogenesis in Helicobacter pylori infection. Gut 43:S2–S5
Shyer AE, Huycke TR, Lee C et al (2015) Bending gradients: how the intestinal stem cell gets its home. Cell 161:569–580. doi:10.1016/j.cell.2015.03.041
Sokolova O, Borgmann M, Rieke C et al (2013) Helicobacter pylori induces type 4 secretion system-dependent, but CagA-independent activation of IκBs and NF-κB/RelA at early time points. Int J Med Microbiol 303:548–552. doi:10.1016/j.ijmm.2013.07.008
Solnick JV, Eaton KA, Peek RM (2016) Animal models of Helicobacter pylori infection. In: Backert S, Yamaoka Y (eds) Helicobacter pylori research: from bench to bedside. Springer, Japan, Tokyo, pp 273–297. doi:10.1242/dmm.000364
Stange DE, Koo B-K, Huch M et al (2013) Differentiated Troy+ chief cells act as reserve stem cells to generate all lineages of the stomach epithelium. Cell 155:357–368. doi:10.1016/j.cell.2013.09.008
Tung Y-C, Hsiao AY, Allen SG et al (2011) High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. The Analyst 136:473–478. doi:10.1039/c0an00609b
Van de Wetering M, Francies HE, Francis JM et al (2015) Prospective derivation of a living organoid biobank of colorectal cancer patients. Cell 161:933–945. doi:10.1016/j.cell.2015.03.053
Van Den Brink GR, Hardwick JCH, Peppelenbosch MP et al (2001) Sonic hedgehog regulates gastric gland morphogenesis in man and mouse. Gastroenterology 121:317–328. doi:10.1053/gast.2001.26261
VanDussen KL, Marinshaw JM, Shaikh N et al (2015) Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays. Gut 64:911–920. doi:10.1136/gutjnl-2013-306651
Wang TC, Koh TJ, Varro A et al (1996) Processing and proliferative effects of human progastrin in transgenic mice. J Clin Invest 98:1918–1929. doi:10.1172/JCI118993
Warren JR, Marshall B (1983) Unidentified curved bacilli on gastric epithelium in active chronic gastritis. The Lancet 321:1273–1275. doi:10.1016/S0140-6736(83)92719-8
Wells JM, Spence JR (2014) How to make an intestine. Development 141:752–760. doi:10.1242/dev.097386
Werner K, Weitz J, Stange DE (2016) Organoids as model systems for gastrointestinal diseases: tissue engineering meets genetic engineering. Curr Pathobiol Rep 4:1–9. doi:10.1007/s40139-016-0100-z
Wroblewski LE, Piazuelo MB, Chaturvedi R et al (2014) Helicobacter pylori targets cancer-associated apical-junctional constituents in gastroids and gastric epithelial cells. Gut. doi:10.1136/gutjnl-2014-307650
Young DF, Carlos TS, Hagmaier K et al (2007) AGS and other tissue culture cells can unknowingly be persistently infected with PIV5; a virus that blocks interferon signalling by degrading STAT1. Virology 365:238–240. doi:10.1016/j.virol.2007.03.061
Yui S, Nakamura T, Sato T et al (2012) Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+ stem cell. Nat Med 18:618–623. doi:10.1038/nm.2695
Acknowledgements
We thank Stan Gorski, Yana Zavros and Jaap Löwenthal for critical reading of the manuscript and constructive comments for improvements.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer International Publishing AG
About this chapter
Cite this chapter
Pompaiah, M., Bartfeld, S. (2017). Gastric Organoids: An Emerging Model System to Study Helicobacter pylori Pathogenesis. In: Tegtmeyer, N., Backert, S. (eds) Molecular Pathogenesis and Signal Transduction by Helicobacter pylori. Current Topics in Microbiology and Immunology, vol 400. Springer, Cham. https://doi.org/10.1007/978-3-319-50520-6_7
Download citation
DOI: https://doi.org/10.1007/978-3-319-50520-6_7
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-50519-0
Online ISBN: 978-3-319-50520-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)