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Isolation and propagation of primary human cholangiocyte organoids for the generation of bioengineered biliary tissue.
- Source :
-
Nature protocols [Nat Protoc] 2019 Jun; Vol. 14 (6), pp. 1884-1925. Date of Electronic Publication: 2019 May 20. - Publication Year :
- 2019
-
Abstract
- Pediatric liver transplantation is often required as a consequence of biliary disorders because of the lack of alternative treatments for repairing or replacing damaged bile ducts. To address the lack of availability of pediatric livers suitable for transplantation, we developed a protocol for generating bioengineered biliary tissue suitable for biliary reconstruction. Our platform allows the derivation of cholangiocyte organoids (COs) expressing key biliary markers and retaining functions of primary extra- or intrahepatic duct cholangiocytes within 2 weeks of isolation. COs are subsequently seeded on polyglycolic acid (PGA) scaffolds or densified collagen constructs for 4 weeks to generate bioengineered tissue retaining biliary characteristics. Expertise in organoid culture and tissue engineering is desirable for optimal results. COs correspond to mature functional cholangiocytes, differentiating our method from alternative organoid systems currently available that propagate adult stem cells. Consequently, COs provide a unique platform for studies in biliary physiology and pathophysiology, and the resulting bioengineered tissue has broad applications for regenerative medicine and cholangiopathies.
- Subjects :
- Animals
Biocompatible Materials chemistry
Cell Separation methods
Cells, Cultured
Equipment Design
Humans
Mice
Tissue Engineering instrumentation
Tissue Scaffolds chemistry
Bile Ducts cytology
Bile Ducts physiology
Organoids cytology
Organoids physiology
Regeneration
Tissue Engineering methods
Subjects
Details
- Language :
- English
- ISSN :
- 1750-2799
- Volume :
- 14
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Nature protocols
- Publication Type :
- Academic Journal
- Accession number :
- 31110298
- Full Text :
- https://doi.org/10.1038/s41596-019-0168-0