Abstract
Intestinal sub-epithelial myofibroblasts (ISEMFs) are mesenchymal cells that do not express cytokeratin but express α-smooth muscle actin and vimentin. Despite being cells with diverse functions, there is a paucity of knowledge about their origin and functions primarily due to the absence of a stable cell line. Although myofibroblast in vitro models for human, mouse, and pig are available, there is no ISEMF cell line available from young calves. We isolated and developed an ileal ISEMF cell line from a 2-d-old calf that expressed α-smooth muscle actin and vimentin but no cytokeratin indicating true myofibroblast cells. To overcome replicative senescence, we immortalized primary cells with SV40 large T antigen. We characterized and compared both primary and immortalized ileal ISEMF cells for surface glycan and Toll-like-receptor (TLR) expression by lectin-binding assay and real-time quantitative PCR (RT-qPCR) assay respectively. SV40 immortalization significantly decreased surface lectin binding for lectins GSL-I, PHA-L, ECL, Jacalin, Con-A, LCA, and LEL. Both cell types expressed TLRs 1–9 and showed no significant differences in TLR expression. Thus, these cells can be useful in vitro model to study ISEMF’s origin, physiology, and functions.
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References
Akira S, Takeda K (2004) Toll-like receptor signalling. Nat Rev Immunol 4(7):499–511. https://doi.org/10.1038/nri1391
Andoh A, Bamba S, Brittan M, Fujiyama Y, Wright NA (2007) Role of intestinal subepithelial myofibroblasts in inflammation and regenerative response in the gut. Pharmacol Ther 114(1):94–106. https://doi.org/10.1016/j.pharmthera.2006.12.004
Audfray A, Claudinon J, Abounit S, Ruvoen-Clouet N, Larson G, Smith DF, Wimmerova M, Le Pendu J, Romer W, Varrot A, Imberty A (2012) Fucose-binding lectin from opportunistic pathogen Burkholderia ambifaria binds to both plant and human oligosaccharidic epitopes. J Biol Chem 287(6):4335–4347. https://doi.org/10.1074/jbc.M111.314831
Begum S, Quach J, Chadee K (2015) Immune evasion mechanisms of Entamoeba histolytica: progression to disease. Front Microbiol 6:1394. https://doi.org/10.3389/fmicb.2015.01394
Bochaton-Piallat ML, Gabbiani G, Hinz B (2016) The myofibroblast in wound healing and fibrosis: answered and unanswered questions. F1000Res 5. https://doi.org/10.12688/f1000research.8190.1
Bochner BS, Zimmermann N (2015) Role of siglecs and related glycan-binding proteins in immune responses and immunoregulation. J Allergy Clin Immunol 135(3):598–608. https://doi.org/10.1016/j.jaci.2014.11.031
Brinchmann MF, Patel DM, Pinto N, Iversen MH (2018) Functional aspects of fish mucosal lectins-interaction with non-self. Molecules 23(5). https://doi.org/10.3390/molecules23051119
Brudner M, Karpel M, Lear C, Chen L, Yantosca LM, Scully C, Sarraju A, Sokolovska A, Zariffard MR, Eisen DP, Mungall BA, Kotton DN, Omari A, Huang IC, Farzan M, Takahashi K, Stuart L, Stahl GL, Ezekowitz AB, Spear GT, Olinger GG, Schmidt EV, Michelow IC (2013) Lectin-dependent enhancement of Ebola virus infection via soluble and transmembrane C-type lectin receptors. PLoS One 8(4):e60838. https://doi.org/10.1371/journal.pone.0060838
Bruyneel EA, Debray H, De Mets M, Mareel MM, Montreuil J (1990) Altered glycosylation in Madin-Darby canine kidney (MDCK) cells after transformation by murine sarcoma virus. Clin Exp Metastasis 8(3):241–253
Cao X (2016) Self-regulation and cross-regulation of pattern-recognition receptor signalling in health and disease. Nat Rev Immunol 16(1):35–50. https://doi.org/10.1038/nri.2015.8
Carmolli M, Duggal P, Haque R, Lindow J, Mondal D, Petri WA Jr, Mourningstar P, Larsson CJ, Sreenivasan M, Khan S, Kirkpatrick BD (2009) Deficient serum mannose-binding lectin levels and MBL2 polymorphisms increase the risk of single and recurrent Cryptosporidium infections in young children. J Infect Dis 200(10):1540–1547. https://doi.org/10.1086/606013
Casanova JL, Abel L (2004) Human mannose-binding lectin in immunity: friend, foe, or both? J Exp Med 199(10):1295–1299. https://doi.org/10.1084/jem.20040537
Cristofalo VJ, Pignolo RJ (1993) Replicative senescence of human fibroblast-like cells in culture. Physiol Rev 73(3):617–638. https://doi.org/10.1152/physrev.1993.73.3.617
Crosby LM, Moore TM, George M, Yoon LW, Easton MJ, Ni H, Morgan KT, DeAngelo AB (2010) Transformation of SV40-immortalized human uroepithelial cells by 3-methylcholanthrene increases IFN- and large T antigen-induced transcripts. Cancer Cell Int 10(1):4. https://doi.org/10.1186/1475-2867-10-4
Dam TK, Brewer CF (2010) Lectins as pattern recognition molecules: the effects of epitope density in innate immunity. Glycobiology 20(3):270–279. https://doi.org/10.1093/glycob/cwp186
Dambuza IM, Brown GD (2015) C-type lectins in immunity: recent developments. Curr Opin Immunol 32:21–27. https://doi.org/10.1016/j.coi.2014.12.002
Darby IA, Laverdet B, Bonte F, Desmouliere A (2014) Fibroblasts and myofibroblasts in wound healing. Clin Cosmet Investig Dermatol 7:301–311. https://doi.org/10.2147/CCID.S50046
Day CJ, Tran EN, Semchenko EA, Tram G, Hartley-Tassell LE, Ng PS, King RM, Ulanovsky R, McAtamney S, Apicella MA, Tiralongo J, Morona R, Korolik V, Jennings MP (2015) Glycan:glycan interactions: high affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells. Proc Natl Acad Sci U S A 112(52):E7266–E7275. https://doi.org/10.1073/pnas.1421082112
De Semir D, Maurisse R, Vock E, Gruenert D (2008) Immortalization strategies for epithelial cells in primary culture. https://doi.org/10.1007/978-0-387-74901-3_26
De Wever O, Demetter P, Mareel M, Bracke M (2008) Stromal myofibroblasts are drivers of invasive cancer growth. Int J Cancer 123(10):2229–2238. https://doi.org/10.1002/ijc.23925
Eun K, Hwang SU, Jeong YW, Seo S, Lee SY, Hwang WS, Hyun SH, Kim H (2017) SV40 large T antigen disrupts embryogenesis of canine and porcine somatic cell nuclear transfer embryo. Biol Proced Online 19(1):13. https://doi.org/10.1186/s12575-017-0061-6
Eyden B (2008) The myofibroblast: phenotypic characterization as a prerequisite to understanding its functions in translational medicine. J Cell Mol Med 12(1):22–37. https://doi.org/10.1111/j.1582-4934.2007.00213.x
Frederick JR, Petri WA Jr (2005) Roles for the galactose-/N-acetylgalactosamine-binding lectin of Entamoeba in parasite virulence and differentiation. Glycobiology 15(12):53R–59R. https://doi.org/10.1093/glycob/cwj007
Fritsch C, Swietlicki EA, Lefebvre O, Kedinger M, Iordanov H, Levin MS, Rubin DC (2002) Epimorphin expression in intestinal myofibroblasts induces epithelial morphogenesis. J Clin Invest 110(11):1629–1641. https://doi.org/10.1172/JCI13588
Furuya K, Sokabe M, Furuya S (2005) Characteristics of subepithelial fibroblasts as a mechano-sensor in the intestine: cell-shape-dependent ATP release and P2Y1 signaling. J Cell Sci 118 (Pt 15:3289–3304. https://doi.org/10.1242/jcs.02453
Fuster MM, Esko JD (2005) The sweet and sour of cancer: glycans as novel therapeutic targets. Nat Rev Cancer 5(7):526–542. https://doi.org/10.1038/nrc1649
Ghufran MS, Ghosh K, Kanade SR (2017) A fucose specific lectin from Aspergillus flavus induced interleukin-8 expression is mediated by mitogen activated protein kinase p38. Med Mycol 55(3):323–333. https://doi.org/10.1093/mmy/myw066
Hashim OH, Jayapalan JJ, Lee CS (2017) Lectins: an effective tool for screening of potential cancer biomarkers. PeerJ 5:e3784. https://doi.org/10.7717/peerj.3784
Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636
Hinz B, Phan SH, Thannickal VJ, Galli A, Bochaton-Piallat ML, Gabbiani G (2007) The myofibroblast: one function, multiple origins. Am J Pathol 170(6):1807–1816. https://doi.org/10.2353/ajpath.2007.070112
Holst S, Belo AI, Giovannetti E, van Die I, Wuhrer M (2017) Profiling of different pancreatic cancer cells used as models for metastatic behaviour shows large variation in their N-glycosylation. Sci Rep 7(1):16623. https://doi.org/10.1038/s41598-017-16811-6
Ibrahim D (2018) Glycoconjugates pattern and chemosensory cells in the camel respiratory mucosa: lectin and immunohistochemical studies. Tissue Cell 51:84–90. https://doi.org/10.1016/j.tice.2018.03.006
Ielasi FS, Alioscha-Perez M, Donohue D, Claes S, Sahli H, Schols D, Willaert RG (2016) Lectin-glycan interaction network-based identification of host receptors of microbial pathogenic adhesins. MBio 7(4). https://doi.org/10.1128/mBio.00584-16
Itakura Y, Sasaki N, Kami D, Gojo S, Umezawa A, Toyoda M (2016) N- and O-glycan cell surface protein modifications associated with cellular senescence and human aging. Cell Biosci 6(1):14. https://doi.org/10.1186/s13578-016-0079-5
Iwanaga K, Murata T, Hori M, Ozaki H (2010) Isolation and characterization of bovine intestinal subepithelial myofibroblasts. J Pharmacol Sci 112(1):98–104. https://doi.org/10.1254/jphs.09258FP
Janovick-Guretzky NA, Dann HM, Carlson DB, Murphy MR, Loor JJ, Drackley JK (2007) Housekeeping gene expression in bovine liver is affected by physiological state, feed intake, and dietary treatment. J Dairy Sci 90(5):2246–2252. https://doi.org/10.3168/jds.2006-640
Katwal P, Thomas M, Uprety T, Hildreth MB, Kaushik RS (2019) Development and biochemical and immunological characterization of early passage and immortalized bovine intestinal epithelial cell lines from the ileum of a young calf. Cytotechnology 71(1):127–148. https://doi.org/10.1007/s10616-018-0272-y
Kaushik RS, Begg AA, Wilson HL, Aich P, Abrahamsen MS, Potter A, Babiuk LA, Griebel P (2008) Establishment of fetal bovine intestinal epithelial cell cultures susceptible to bovine rotavirus infection. J Virol Methods 148(1–2):182–196. https://doi.org/10.1016/j.jviromet.2007.11.006
Kawasaki H, Ohama T, Hori M, Sato K (2013) Establishment of mouse intestinal myofibroblast cell lines. World J Gastroenterol 19(17):2629–2637. https://doi.org/10.3748/wjg.v19.i17.2629
Kaye GI, Lane N, Pascal RR (1968) Colonic pericryptal fibroblast sheath: replication, migration, and cytodifferentiation of a mesenchymal cell system in adult tissue. II. Fine structural aspects of normal rabbit and human colon. Gastroenterology 54(5):852–865
Kim YJ, Varki A (1997) Perspectives on the significance of altered glycosylation of glycoproteins in cancer. Glycoconj J 14(5):569–576. https://doi.org/10.1023/a:1018580324971
Lahar N, Lei NY, Wang J, Jabaji Z, Tung SC, Joshi V, Lewis M, Stelzner M, Martin MG, Dunn JC (2011) Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium. PLoS One 6(11):e26898. https://doi.org/10.1371/journal.pone.0026898
Lei NY, Jabaji Z, Wang J, Joshi VS, Brinkley GJ, Khalil H, Wang F, Jaroszewicz A, Pellegrini M, Li L, Lewis M, Stelzner M, Dunn JC, Martin MG (2014) Intestinal subepithelial myofibroblasts support the growth of intestinal epithelial stem cells. PLoS One 9(1):e84651. https://doi.org/10.1371/journal.pone.0084651
Lekka M, Laidler P, Labedz M, Kulik AJ, Lekki J, Zajac W, Stachura Z (2006) Specific detection of glycans on a plasma membrane of living cells with atomic force microscopy. Chem Biol 13(5):505–512. https://doi.org/10.1016/j.chembiol.2006.03.006
Levine AJ (2009) The common mechanisms of transformation by the small DNA tumor viruses: the inactivation of tumor suppressor gene products: p53. Virology 384(2):285–293. https://doi.org/10.1016/j.virol.2008.09.034
Lis H, Sharon N (1986) Lectins as molecules and as tools. Annu Rev Biochem 55:35–67. https://doi.org/10.1146/annurev.bi.55.070186.000343
Liu X, Nie H, Zhang Y, Yao Y, Maitikabili A, Qu Y, Shi S, Chen C, Li Y (2013) Cell surface-specific N-glycan profiling in breast cancer. PLoS One 8(8):e72704. https://doi.org/10.1371/journal.pone.0072704
Majno G, Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR (1971) Contraction of granulation tissue in vitro: similarity to smooth muscle. Science 173(3996):548–550
Mifflin RC, Pinchuk IV, Saada JI, Powell DW (2011) Intestinal myofibroblasts: targets for stem cell therapy. Am J Physiol Gastrointest Liver Physiol 300(5):G684–G696. https://doi.org/10.1152/ajpgi.00474.2010
Nizet V, Varki A, Aebi M (2015) Microbial lectins: hemagglutinins, adhesins, and toxins. In: Varki A, Cummings RD et al (eds) Essentials of Glycobiology. Cold Spring Harbor (NY), pp 481–491. https://doi.org/10.1101/glycobiology.3e.037
Ohama T, Okada M, Murata T, Brautigan DL, Hori M, Ozaki H (2008) Sphingosine-1-phosphate enhances IL-1{beta}-induced COX-2 expression in mouse intestinal subepithelial myofibroblasts. Am J Physiol Gastrointest Liver Physiol 295(4):G766–G775. https://doi.org/10.1152/ajpgi.90423.2008
Otte JM, Rosenberg IM, Podolsky DK (2003) Intestinal myofibroblasts in innate immune responses of the intestine. Gastroenterology 124(7):1866–1878. https://doi.org/10.1016/s0016-5085(03)00403-7
Owens BM (2015) Inflammation, innate immunity, and the intestinal stromal cell niche: opportunities and challenges. Front Immunol 6(319):319. https://doi.org/10.3389/fimmu.2015.00319
Padler-Karavani V (2014) Aiming at the sweet side of cancer: aberrant glycosylation as possible target for personalized-medicine. Cancer Lett 352(1):102–112. https://doi.org/10.1016/j.canlet.2013.10.005
Pinchuk IV, Mifflin RC, Saada JI, Powell DW (2010) Intestinal mesenchymal cells. Curr Gastroenterol Rep 12(5):310–318. https://doi.org/10.1007/s11894-010-0135-y
Pipas JM (2009) SV40: cell transformation and tumorigenesis. Virology 384(2):294–303. https://doi.org/10.1016/j.virol.2008.11.024
Pourreyron C, Dumortier J, Ratineau C, Nejjari M, Beatrix O, Jacquier MF, Remy L, Chayvialle JA, Scoazec JY (2003) Age-dependent variations of human and rat colon myofibroblasts in culture: influence on their functional interactions with colon cancer cells. Int J Cancer 104(1):28–35. https://doi.org/10.1002/ijc.10898
Powell DW, Mifflin RC, Valentich JD, Crowe SE, Saada JI, West AB (1999a) Myofibroblasts. I. Paracrine cells important in health and disease. Am J Phys 277(1 Pt 1):C1–C9
Powell DW, Mifflin RC, Valentich JD, Crowe SE, Saada JI, West AB (1999b) Myofibroblasts. II. Intestinal subepithelial myofibroblasts. Am J Phys 277(2):C183–C201. https://doi.org/10.1152/ajpcell.1999.277.2.C183
Powell DW, Pinchuk IV, Saada JI, Chen X, Mifflin RC (2011) Mesenchymal cells of the intestinal lamina propria. Annu Rev Physiol 73:213–237. https://doi.org/10.1146/annurev.physiol.70.113006.100646
Przybylo M, Litynska A, Pochec E (2005) Different adhesion and migration properties of human HCV29 non-malignant urothelial and T24 bladder cancer cells: role of glycosylation. Biochimie 87(2):133–142. https://doi.org/10.1016/j.biochi.2004.12.003
Pucilowska JB, McNaughton KK, Mohapatra NK, Hoyt EC, Zimmermann EM, Sartor RB, Lund PK (2000) IGF-I and procollagen alpha1(I) are coexpressed in a subset of mesenchymal cells in active Crohn's disease. Am J Physiol Gastrointest Liver Physiol 279(6):G1307–G1322. https://doi.org/10.1152/ajpgi.2000.279.6.G1307
Rathi AV, Saenz Robles MT, Cantalupo PG, Whitehead RH, Pipas JM (2009) Simian virus 40 T-antigen-mediated gene regulation in enterocytes is controlled primarily by the Rb-E2F pathway. J Virol 83(18):9521–9531. https://doi.org/10.1128/JVI.00583-09
Rieder F, Fiocchi C (2008) Intestinal fibrosis in inflammatory bowel disease - current knowledge and future perspectives. J Crohns Colitis 2(4):279–290. https://doi.org/10.1016/j.crohns.2008.05.009
Rosbjerg A, Genster N, Pilely K, Garred P (2017) Evasion mechanisms used by pathogens to escape the lectin complement pathway. Front Microbiol 8(868):868. https://doi.org/10.3389/fmicb.2017.00868
Rostand KS, Esko JD (1997) Microbial adherence to and invasion through proteoglycans. Infect Immun 65(1):1–8
Roulis M, Flavell RA (2016) Fibroblasts and myofibroblasts of the intestinal lamina propria in physiology and disease. Differentiation 92(3):116–131. https://doi.org/10.1016/j.diff.2016.05.002
Saada JI, Pinchuk IV, Barrera CA, Adegboyega PA, Suarez G, Mifflin RC, Di Mari JF, Reyes VE, Powell DW (2006) Subepithelial myofibroblasts are novel nonprofessional APCs in the human colonic mucosa. J Immunol 177(9):5968–5979. https://doi.org/10.4049/jimmunol.177.9.5968
Saenz Robles MT, Shivalila C, Wano J, Sorrells S, Roos A, Pipas JM (2013) Two independent regions of simian virus 40 T antigen increase CBP/p300 levels, alter patterns of cellular histone acetylation, and immortalize primary cells. J Virol 87(24):13499–13509. https://doi.org/10.1128/JVI.02658-13
Sasaki N, Itakura Y, Toyoda M (2017) Sialylation regulates myofibroblast differentiation of human skin fibroblasts. Stem Cell Res Ther 8(1):81. https://doi.org/10.1186/s13287-017-0534-1
Shao J, Sheng H (2010) Amphiregulin promotes intestinal epithelial regeneration: roles of intestinal subepithelial myofibroblasts. Endocrinology 151(8):3728–3737. https://doi.org/10.1210/en.2010-0319
Simmons JG, Pucilowska JB, Keku TO, Lund PK (2002) IGF-I and TGF-beta1 have distinct effects on phenotype and proliferation of intestinal fibroblasts. Am J Physiol Gastrointest Liver Physiol 283(3):G809–G818. https://doi.org/10.1152/ajpgi.00057.2002
Skalli O, Schurch W, Seemayer T, Lagace R, Montandon D, Pittet B, Gabbiani G (1989) Myofibroblasts from diverse pathologic settings are heterogeneous in their content of actin isoforms and intermediate filament proteins. Lab Investig 60(2):275–285
Song X, Jin P, Qin S, Chen L, Ma F (2012) The evolution and origin of animal toll-like receptor signaling pathway revealed by network-level molecular evolutionary analyses. PLoS One 7(12):e51657. https://doi.org/10.1371/journal.pone.0051657
Stowell SR, Ju T, Cummings RD (2015) Protein glycosylation in cancer. Annu Rev Pathol 10:473–510. https://doi.org/10.1146/annurev-pathol-012414-040438
Takanashi N, Tomosada Y, Villena J, Murata K, Takahashi T, Chiba E, Tohno M, Shimazu T, Aso H, Suda Y, Ikegami S, Itoh H, Kawai Y, Saito T, Alvarez S, Kitazawa H (2013) Advanced application of bovine intestinal epithelial cell line for evaluating regulatory effect of lactobacilli against heat-killed enterotoxigenic Escherichia coli-mediated inflammation. BMC Microbiol 13(1):54. https://doi.org/10.1186/1471-2180-13-54
Thaiss CA, Levy M, Itav S, Elinav E (2016) Integration of innate immune signaling. Trends Immunol 37(2):84–101. https://doi.org/10.1016/j.it.2015.12.003
Thomas M, Pierson M, Uprety T, Zhu L, Ran Z, Sreenivasan CC, Wang D, Hause B, Francis DH, Li F, Kaushik RS (2018) Comparison of porcine airway and intestinal epithelial cell lines for the susceptibility and expression of pattern recognition receptors upon influenza virus infection. Viruses 10(6). https://doi.org/10.3390/v10060312
Tytgat HLP, de Vos WM (2016) Sugar coating the envelope: glycoconjugates for microbe-host crosstalk. Trends Microbiol 24(11):853–861. https://doi.org/10.1016/j.tim.2016.06.004
Valatas V, Filidou E, Drygiannakis I, Kolios G (2017) Stromal and immune cells in gut fibrosis: the myofibroblast and the scarface. Ann Gastroenterol 30(4):393–404. https://doi.org/10.20524/aog.2017.0146
Valentich JD, Popov V, Saada JI, Powell DW (1997) Phenotypic characterization of an intestinal subepithelial myofibroblast cell line. Am J Phys 272(5 Pt 1):C1513–C1524. https://doi.org/10.1152/ajpcell.1997.272.5.C1513
Walton KL, Holt L, Sartor RB (2009) Lipopolysaccharide activates innate immune responses in murine intestinal myofibroblasts through multiple signaling pathways. Am J Physiol Gastrointest Liver Physiol 296(3):G601–G611. https://doi.org/10.1152/ajpgi.00022.2008
Wasik BR, Barnard KN, Parrish CR (2016) Effects of sialic acid modifications on virus binding and infection. Trends Microbiol 24(12):991–1001. https://doi.org/10.1016/j.tim.2016.07.005
Weis W, Brown JH, Cusack S, Paulson JC, Skehel JJ, Wiley DC (1988) Structure of the influenza virus haemagglutinin complexed with its receptor, sialic acid. Nature 333(6172):426–431. https://doi.org/10.1038/333426a0
Wu LA, Feng J, Wang L, Mu YD, Baker A, Donly KJ, Harris SE, MacDougall M, Chen S (2011) Development and characterization of a mouse floxed Bmp2 osteoblast cell line that retains osteoblast genotype and phenotype. Cell Tissue Res 343(3):545–558. https://doi.org/10.1007/s00441-010-1120-3
Yoshikawa T, Hamada S, Otsuji E, Tsujimoto H, Hagiwara A (2011) Endocrine differentiation of rat enterocytes in long-term three-dimensional co-culture with intestinal myofibroblasts. In Vitro Cell Dev Biol Anim 47(10):707–715. https://doi.org/10.1007/s11626-011-9458-8
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Radhey S. Kaushik and Tirth Uprety were funded and supported by USDA, NIFA, and SDSU Agricultural Experiment Station Hatch grant number SD00H547-15. We acknowledge the use of the SDSU-FGCF supported in part by NSF/EPSCoR grant number 0091948 and by the State of South Dakota.
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This research project was funded and supported by USDA, NIFA, and SDSU Agricultural Experiment Station Hatch grant number SD00H547-15. This study was also funded by USDA/CSREES through the project title JDIP: Johne’s Disease Integrated Program in Research, Education, and Extension, grant number 2004-35605-14243, sub-award No. Q6286224171.
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RSK conceived and designed the study, received funding, and conducted some of the experiments. RSK also reviewed the manuscript. TU, LA, and CS conducted the experiments, analyzed the results, and wrote the paper. BBS is an undergraduate student who helped with lectin profiling of ISEMF cells. AY helped on flow cytometry–based lectin-binding assays. FL and MH contributed to reagents/materials and analysis and helped in microscopy.
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Uprety, T., Spurlin, B.B., Antony, L. et al. Development and characterization of a stable bovine intestinal sub-epithelial myofibroblast cell line from ileum of a young calf. In Vitro Cell.Dev.Biol.-Animal 55, 533–547 (2019). https://doi.org/10.1007/s11626-019-00365-0
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DOI: https://doi.org/10.1007/s11626-019-00365-0