Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns
- Author
- Jennifer Barrila, Jiseon Yang, Aurélie Crabbé (UGent) , Shameema F Sarker, Yulong Liu, C Mark Ott, Mayra A Nelman-Gonzalez, Simon J Clemett, Seth D Nydam, Rebecca J Forsyth, Richard R Davis, Brian E Crucian, Heather Quiriarte, Kenneth L Roland, Karen Brenneman, Clarence Sams, Christine Loscher and Cheryl A Nickerson
- Organization
- Abstract
- Three-dimensional models of human intestinal epithelium mimic the differentiated form and function of parental tissues often not exhibited by two-dimensional monolayers and respond to Salmonella in key ways that reflect in vivo infections. To further enhance the physiological relevance of three-dimensional models to more closely approximate in vivo intestinal microenvironments encountered by Salmonella, we developed and validated a novel three-dimensional co-culture infection model of colonic epithelial cells and macrophages using the NASA Rotating Wall Vessel bioreactor. First, U937 cells were activated upon collagen-coated scaffolds. HT-29 epithelial cells were then added and the three-dimensional model was cultured in the bioreactor until optimal differentiation was reached, as assessed by immunohistochemical profiling and bead uptake assays. The new co-culture model exhibited in vivo-like structural and phenotypic characteristics, including three-dimensional architecture, apical-basolateral polarity, well-formed tight/adherens junctions, mucin, multiple epithelial cell types, and functional macrophages. Phagocytic activity of macrophages was confirmed by uptake of inert, bacteria-sized beads. Contribution of macrophages to infection was assessed by colonization studies of Salmonella pathovars with different host adaptations and disease phenotypes (Typhimurium ST19 strain SL1344 and ST313 strain D23580; Typhi Ty2). In addition, Salmonella were cultured aerobically or microaerobically, recapitulating environments encountered prior to and during intestinal infection, respectively. All Salmonella strains exhibited decreased colonization in co-culture (HT-29-U937) relative to epithelial (HT-29) models, indicating antimicrobial function of macrophages. Interestingly, D23580 exhibited enhanced replication/survival in both models following invasion. Pathovar-specific differences in colonization and intracellular co-localization patterns were observed. These findings emphasize the power of incorporating a series of related three-dimensional models within a study to identify microenvironmental factors important for regulating infection.
- Keywords
- TRANSMEMBRANE CONDUCTANCE REGULATOR, SEROVAR GALLINARUM REQUIRES, COMPLETE GENOME SEQUENCE, STG FIMBRIAL OPERON, IN-VITRO, TYPHIMURIUM INVASION, BACTERIAL SURVIVAL, HOST-CELLS, IVB PILI, TYPHI
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8525928
- MLA
- Barrila, Jennifer, et al. “Three-Dimensional Organotypic Co-Culture Model of Intestinal Epithelial Cells and Macrophages to Study Salmonella Enterica Colonization Patterns.” NPJ MICROGRAVITY, vol. 3, 2017, doi:10.1038/s41526-017-0011-2.
- APA
- Barrila, J., Yang, J., Crabbé, A., Sarker, S. F., Liu, Y., Ott, C. M., … Nickerson, C. A. (2017). Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns. NPJ MICROGRAVITY, 3. https://doi.org/10.1038/s41526-017-0011-2
- Chicago author-date
- Barrila, Jennifer, Jiseon Yang, Aurélie Crabbé, Shameema F Sarker, Yulong Liu, C Mark Ott, Mayra A Nelman-Gonzalez, et al. 2017. “Three-Dimensional Organotypic Co-Culture Model of Intestinal Epithelial Cells and Macrophages to Study Salmonella Enterica Colonization Patterns.” NPJ MICROGRAVITY 3. https://doi.org/10.1038/s41526-017-0011-2.
- Chicago author-date (all authors)
- Barrila, Jennifer, Jiseon Yang, Aurélie Crabbé, Shameema F Sarker, Yulong Liu, C Mark Ott, Mayra A Nelman-Gonzalez, Simon J Clemett, Seth D Nydam, Rebecca J Forsyth, Richard R Davis, Brian E Crucian, Heather Quiriarte, Kenneth L Roland, Karen Brenneman, Clarence Sams, Christine Loscher, and Cheryl A Nickerson. 2017. “Three-Dimensional Organotypic Co-Culture Model of Intestinal Epithelial Cells and Macrophages to Study Salmonella Enterica Colonization Patterns.” NPJ MICROGRAVITY 3. doi:10.1038/s41526-017-0011-2.
- Vancouver
- 1.Barrila J, Yang J, Crabbé A, Sarker SF, Liu Y, Ott CM, et al. Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns. NPJ MICROGRAVITY. 2017;3.
- IEEE
- [1]J. Barrila et al., “Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns,” NPJ MICROGRAVITY, vol. 3, 2017.
@article{8525928, abstract = {{Three-dimensional models of human intestinal epithelium mimic the differentiated form and function of parental tissues often not exhibited by two-dimensional monolayers and respond to Salmonella in key ways that reflect in vivo infections. To further enhance the physiological relevance of three-dimensional models to more closely approximate in vivo intestinal microenvironments encountered by Salmonella, we developed and validated a novel three-dimensional co-culture infection model of colonic epithelial cells and macrophages using the NASA Rotating Wall Vessel bioreactor. First, U937 cells were activated upon collagen-coated scaffolds. HT-29 epithelial cells were then added and the three-dimensional model was cultured in the bioreactor until optimal differentiation was reached, as assessed by immunohistochemical profiling and bead uptake assays. The new co-culture model exhibited in vivo-like structural and phenotypic characteristics, including three-dimensional architecture, apical-basolateral polarity, well-formed tight/adherens junctions, mucin, multiple epithelial cell types, and functional macrophages. Phagocytic activity of macrophages was confirmed by uptake of inert, bacteria-sized beads. Contribution of macrophages to infection was assessed by colonization studies of Salmonella pathovars with different host adaptations and disease phenotypes (Typhimurium ST19 strain SL1344 and ST313 strain D23580; Typhi Ty2). In addition, Salmonella were cultured aerobically or microaerobically, recapitulating environments encountered prior to and during intestinal infection, respectively. All Salmonella strains exhibited decreased colonization in co-culture (HT-29-U937) relative to epithelial (HT-29) models, indicating antimicrobial function of macrophages. Interestingly, D23580 exhibited enhanced replication/survival in both models following invasion. Pathovar-specific differences in colonization and intracellular co-localization patterns were observed. These findings emphasize the power of incorporating a series of related three-dimensional models within a study to identify microenvironmental factors important for regulating infection.}}, articleno = {{10}}, author = {{Barrila, Jennifer and Yang, Jiseon and Crabbé, Aurélie and Sarker, Shameema F and Liu, Yulong and Ott, C Mark and Nelman-Gonzalez, Mayra A and Clemett, Simon J and Nydam, Seth D and Forsyth, Rebecca J and Davis, Richard R and Crucian, Brian E and Quiriarte, Heather and Roland, Kenneth L and Brenneman, Karen and Sams, Clarence and Loscher, Christine and Nickerson, Cheryl A}}, issn = {{2373-8065}}, journal = {{NPJ MICROGRAVITY}}, keywords = {{TRANSMEMBRANE CONDUCTANCE REGULATOR,SEROVAR GALLINARUM REQUIRES,COMPLETE GENOME SEQUENCE,STG FIMBRIAL OPERON,IN-VITRO,TYPHIMURIUM INVASION,BACTERIAL SURVIVAL,HOST-CELLS,IVB PILI,TYPHI}}, language = {{eng}}, pages = {{12}}, title = {{Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns}}, url = {{http://doi.org/10.1038/s41526-017-0011-2}}, volume = {{3}}, year = {{2017}}, }
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