Long-Term Characteristics of Human-Derived Biliary Organoids under a Single Continuous Culture Condition
Abstract
:1. Introduction
2. Materials and Methods
2.1. Organoid Culture
2.2. Immunohistochemistry
2.3. Microscopy
2.4. Single-Nucleus Transcriptome Analysis
3. Results
3.1. Chol-Orgs Were Established from Patients with a Variety of Liver Disorders
3.2. Microscopic Analysis of Chol-Orgs Revealed the Formation of Complex Epithelial Structures with Compartmentalization over Time
3.3. Organoid Immunostaining Revealed Specific Differences between Early-Stage and Late-Stage Organoids
3.4. Electron Microscopic Analysis of Organoids Revealed the Development of Fine Epithelial Cellular Ultrastructure
3.5. Transcriptome Analysis Demonstrates Cholangiocytic Gene Signature of Chol-Orgs
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Patient | Diagnosis | Age (Years) | Organoid Establishment |
---|---|---|---|
1 | Biliary atresia | 1 | No |
2 | Biliary atresia | 0.9 | No |
3 | Maple syrup urine disease | 11 | No |
4 | Progressive familial intrahepatic cholestasis III | 9 | Yes |
6 | Autoimmune hepatitis | 13 | Yes |
7 | Alpha-1 antitrypsin deficiency | 1 | Yes |
8a | Normal liver tissue (hepatoblastoma) | 11 | No |
8b | Hepatoblastoma | 11 | Yes |
9 | Biliary atresia | 1.5 | Yes |
10 | Progressive familial intrahepatic cholestasis III | 4 | Yes |
11 | Ornithine transcarbamylase deficiency | 2 | Yes |
12 | Normal liver tissue (hepatoblastoma) | 3.5 | Yes |
13 | Fibrolamellar carcinoma | 14 | No |
14 | Congenital glycosylation type 1d, mannosyltransferase 6 deficiency | 2.5 | Yes |
15 | Progressive familial intrahepatic cholestasis II | 6 | Yes |
16 | Fibrolamellar carcinoma | 15 | Yes |
Cluster IDs | 2D Chol-Org | 3D Chol-Org |
---|---|---|
1 | 785 (26.78%) | 778 (24.03%) |
2 | 204 (6.96%) | 1018 (31.45%) |
3 | 412 (14.06%) | 426 (13.16%) |
4 | 475 (16.21%) | 348 (10.75%) |
5 | 598 (20.4%) | 76 (2.35%) |
6 | 101 (3.45%) | 320 (9.89%) |
7 | 23 (0.78%) | 245 (7.57%) |
8 | 159 (5.42%) | 15 (0.46%) |
9 | 165 (5.63%) | 6 (0.19%) |
10 | 9 (0.31%) | 5 (0.15%) |
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Aktas, R.G.; Karski, M.; Issac, B.; Sun, L.; Rockowitz, S.; Sliz, P.; Vakili, K. Long-Term Characteristics of Human-Derived Biliary Organoids under a Single Continuous Culture Condition. Cells 2022, 11, 3797. https://doi.org/10.3390/cells11233797
Aktas RG, Karski M, Issac B, Sun L, Rockowitz S, Sliz P, Vakili K. Long-Term Characteristics of Human-Derived Biliary Organoids under a Single Continuous Culture Condition. Cells. 2022; 11(23):3797. https://doi.org/10.3390/cells11233797
Chicago/Turabian StyleAktas, Ranan G., Michael Karski, Biju Issac, Liang Sun, Shira Rockowitz, Piotr Sliz, and Khashayar Vakili. 2022. "Long-Term Characteristics of Human-Derived Biliary Organoids under a Single Continuous Culture Condition" Cells 11, no. 23: 3797. https://doi.org/10.3390/cells11233797