PMID- 29204504 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220408 IS - 2352-345X (Print) IS - 2352-345X (Electronic) IS - 2352-345X (Linking) VI - 4 IP - 1 DP - 2017 Jul TI - Self-renewing Monolayer of Primary Colonic or Rectal Epithelial Cells. PG - 165-182.e7 LID - 10.1016/j.jcmgh.2017.02.011 [doi] AB - BACKGROUND & AIMS: Three-dimensional organoid culture has fundamentally changed the in vitro study of intestinal biology enabling novel assays; however, its use is limited because of an inaccessible luminal compartment and challenges to data gathering in a three-dimensional hydrogel matrix. Long-lived, self-renewing 2-dimensional (2-D) tissue cultured from primary colon cells has not been accomplished. METHODS: The surface matrix and chemical factors that sustain 2-D mouse colonic and human rectal epithelial cell monolayers with cell repertoires comparable to that in vivo were identified. RESULTS: The monolayers formed organoids or colonoids when placed in standard Matrigel culture. As with the colonoids, the monolayers exhibited compartmentalization of proliferative and differentiated cells, with proliferative cells located near the peripheral edges of growing monolayers and differentiated cells predominated in the central regions. Screening of 77 dietary compounds and metabolites revealed altered proliferation or differentiation of the murine colonic epithelium. When exposed to a subset of the compound library, murine organoids exhibited similar responses to that of the monolayer but with differences that were likely attributable to the inaccessible organoid lumen. The response of the human primary epithelium to a compound subset was distinct from that of both the murine primary epithelium and human tumor cells. CONCLUSIONS: This study demonstrates that a self-renewing 2-D murine and human monolayer derived from primary cells can serve as a physiologically relevant assay system for study of stem cell renewal and differentiation and for compound screening. The platform holds transformative potential for personalized and precision medicine and can be applied to emerging areas of disease modeling and microbiome studies. FAU - Wang, Yuli AU - Wang Y AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - DiSalvo, Matthew AU - DiSalvo M AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. FAU - Gunasekara, Dulan B AU - Gunasekara DB AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - Dutton, Johanna AU - Dutton J AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. FAU - Proctor, Angela AU - Proctor A AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - Lebhar, Michael S AU - Lebhar MS AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. FAU - Williamson, Ian A AU - Williamson IA AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. FAU - Speer, Jennifer AU - Speer J AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - Howard, Riley L AU - Howard RL AD - Department of Applied Physical Sciences, University of North Carolina, Chapel Hill, North Carolina. FAU - Smiddy, Nicole M AU - Smiddy NM AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - Bultman, Scott J AU - Bultman SJ AD - Department of Genetics, University of North Carolina, Chapel Hill, North Carolina. FAU - Sims, Christopher E AU - Sims CE AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. FAU - Magness, Scott T AU - Magness ST AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. FAU - Allbritton, Nancy L AU - Allbritton NL AD - Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina. AD - Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, and North Carolina State University, Raleigh, North Carolina. AD - Department of Applied Physical Sciences, University of North Carolina, Chapel Hill, North Carolina. LA - eng GR - R01 CA177993/CA/NCI NIH HHS/United States GR - R01 DK109559/DK/NIDDK NIH HHS/United States PT - Journal Article DEP - 20170306 PL - United States TA - Cell Mol Gastroenterol Hepatol JT - Cellular and molecular gastroenterology and hepatology JID - 101648302 PMC - PMC5710741 OTO - NOTNLM OT - 2-D, two-dimensional OT - 3-D, three-dimensional OT - ALP, alkaline phosphatase OT - CAG, cytomegalovirus enhancer plus chicken actin promoter OT - CI, confidence interval OT - Colonic Epithelial Cells OT - Compound Screening OT - ECM, extracellular matrix OT - EDU, 5-ethynyl-2'-deoxyuridine OT - EGF, epidermal growth factor OT - ENR-W, cell medium with [Wnt-3A] of 30 ng/mL OT - ENR-w, cell medium with [Wnt-3A] of 10 ng/mL OT - HISC, human intestinal stem cell medium OT - IACUC, Institutional Animal Care and Use Committee OT - ISC, intestinal stem cell OT - Monolayer OT - Organoids OT - PBS, phosphate-buffered saline OT - PDMS, polydimethylsiloxane OT - RFP, red fluorescent protein OT - SEM, scanning electron microscope OT - SSMD, strictly standardized mean difference OT - UNC, University of North Carolina OT - alpha-ChgA, anti-chromogranin A OT - alpha-Muc2, anti-mucin2 EDAT- 2017/12/06 06:00 MHDA- 2017/12/06 06:01 PMCR- 2017/03/06 CRDT- 2017/12/06 06:00 PHST- 2016/08/25 00:00 [received] PHST- 2017/02/15 00:00 [accepted] PHST- 2017/12/06 06:00 [entrez] PHST- 2017/12/06 06:00 [pubmed] PHST- 2017/12/06 06:01 [medline] PHST- 2017/03/06 00:00 [pmc-release] AID - S2352-345X(17)30046-2 [pii] AID - 10.1016/j.jcmgh.2017.02.011 [doi] PST - epublish SO - Cell Mol Gastroenterol Hepatol. 2017 Mar 6;4(1):165-182.e7. doi: 10.1016/j.jcmgh.2017.02.011. eCollection 2017 Jul.