# Modeling Epithelial Homeostasis and Perturbation in Three-Dimensional Human Esophageal Organoids
## 5 Conclusions  
In summary, we have established a HOME0 medium formulation and validated its utility to grow single-cell-derived normal human esophageal organoids, enabling the comparison of species differences in esophageal epithelial characteristics as well as organoids grown in the broadly utilized ADF-based medium supplemented with different factors and agents.  
**Supplementary Materials:** The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/biom14091126/s1](https://www.mdpi.com/article/10.3390/biom14091126/s1)]([https://www.mdpi.com/article/10.3390/biom14091126/s1](https://www.mdpi.com/article/10.3390/biom14091126/s1)), Figure S1: Pharmacological EGFR inhibition results in organoid cell death in the absence of exogenous EGF; Figure S2: Normal esophageal PDOs display limited replicative lifespan; Figure S3: A representative image of Halo AI usage for organoid tissue segmentation and analysis of IHC; Figure S4: UAB30 induction of HB-EGF and EGFR tyrosine phosphorylation.  
**Author Contributions:** Conceptualization, H.N., F.M.-H. and K.I.W.; methodology, M.S., C.M. and F.M.-H.; formal analysis, M.S., M.M., W.H., Y.T., N.M., S.F., A.S.P., U.M.S., F.M.-H. and A.J.K.-S.; resources, B.R.W., F.G.O., V.R.A., C.A.E., J.A.A., A.B.M. and F.M.-H.; data curation, M.S., M.M., W.H., Y.T., N.M., E.H.O., M.S.E., C.Y.L. and W.R.B.; writing--original draft preparation, M.S., M.M. and H.N.; writing--review and editing, S.F., U.M.S. and C.M.; project administration, H.N.; funding acquisition, F.M.-H., K.I.W. and H.N. All authors have read and agreed to the published version of the manuscript.  
**Funding:** This study was supported by NIH F01 CA098101 (H.N.), NIH R01 DK114436 (H.N.), NIH R01 AA026297 (H.N.), NIH I30 CA264714 (S.F), and NIH I732 CA265828-01A1 (S.F). [PERSON] PhD was supported by a Postdoctoral Research Fellowship, PF-23-1151788-01-DMC, from the American Cancer Society, [[https://doi.org/10.53354/ACS.PF-23-1151788-01-DMC.pc.gr.175436](https://doi.org/10.53354/ACS.PF-23-1151788-01-DMC.pc.gr.175436)]([https://doi.org/10.53354/ACS.PF-23-1151788-01-DMC.pc.gr.175436](https://doi.org/10.53354/ACS.PF-23-1151788-01-DMC.pc.gr.175436)) (accessed on 1 January 2024). Further, these studies used the resources of the Herbert Irving Comprehensive Cancer Center Molecular Pathology Shared Resource funded in part through Center Grant P30 CA013696 and Organoid & Cell Culture Core of the Columbia University Digestive and Liver Diseases Research Center funded in part through Center Grant P30 DK132710. H.N., F.G.O., and K.I.W. are the recipients of the Fanconi Cancer Foundation grant.  
**Institutional Review Board Statement:** The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Boards of Columbia University (IRB-AAAS4603, latest approval date 23 December 2023) and Children's Hospital of Philadelphia (IRB 13-010552, latest approval date 27 February 2024) for studies involving humans.  
**Informed Consent Statement:** Informed consent was obtained from all the subjects involved in the study.  
**Data Availability Statement:** The original contributions presented in the study are included in the article/Supplementary Materials; further inquiries can be directed to the corresponding author.  
**Acknowledgments:** We thank [PERSON] (NCI P01 Mechanisms of Esophageal Carcinogenesis) and members of the [PERSON] and [PERSON] lab for helpful discussions. We thank the Shared Resources (Flow Cytometry, Molecular Pathology, and Confocal & Specialized Microscopy) at the Herbert Irving Comprehensive Cancer Center at Columbia University for technical support.