Abstract
The biliary tree is an essential component of transplantable human liver tissue. Despite recent advances in liver tissue engineering, attempts at re-creating the intrahepatic biliary tree have not progressed significantly. The finer branches of the biliary tree are structurally and functionally complex and heterogeneous and require harnessing innate developmental processes for their regrowth. Here we demonstrate the ability of decellularized liver extracellular matrix (dECM) hydrogels to induce the in vitro formation of complex biliary networks using encapsulated immortalized mouse small biliary epithelial cells (cholangiocytes). This phenomenon is not observed using immortalized mouse large cholangiocytes, or with purified collagen 1 gels or Matrigel. We also show phenotypic stability via immunostaining for specific cholangiocyte markers. Moreover, tight junction formation and maturation was observed to occur between cholangiocytes, exhibiting polarization and transporter activity. To better define the mechanism of duct formation, we utilized three fluorescently labeled, but otherwise identical populations of cholangiocytes. The cells, in a proximity dependent manner, either branch out clonally, radiating from a single nucleation point, or assemble into multi-colored structures arising from separate populations. These findings present liver dECM as a promising biomaterial for intrahepatic bile duct tissue engineering and as a tool to study duct remodeling in vitro.
Original language | English (US) |
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Article number | 12220 |
Journal | Scientific reports |
Volume | 8 |
Issue number | 1 |
DOIs | |
State | Published - Dec 1 2018 |
Funding
The authors would like to thank Dr. Alexandra Rutz for the development and optimization of decellularization and gel formation protocols. The authors would also like to thank Lennell Reynolds for TEM processing. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. Cytation 3 Plate reader usage and Anton Parr MCR302 Rheometer usage was performed in the Analytical BioNanoTechnology Core Facility of the Simpson Querrey Institute (SQI) at Northwestern University. The U.S. Army Research Office, the U.S. Army Medical Research and Materiel Command, and Northwestern University provided funding to develop SQI and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205). This work was supported by the Northwestern University – Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI CA060553). Flow Cytometry Cell Sorting was performed on a BD FACSAria SORP system, purchased through the support of NIH 1S10OD011996-01. This work was supported by the National Institutes of Health grant number 1K01DK099454, Primary Sclerosing Cholangitis (PSC) Partners Seeking a Cure grant number 7280909288FF, and Northwestern University’s Biotechnology Training Program Cluster Award. This work was supported by a National Institutes of Health Predoctoral Biotechnology Training Program (NIGMS T32 GM008449) and a Ruth L. Kirschstein National Research Service Award Individual Predoctoral Fellowship (NRSA F31 NIDDK 1F31DK108544-01A1) awarded to J.S. Portion of this work was supported in part by the Dr. Nicholas C. Hightower Centennial Chair of Gastroenterology from Scott & White, a VA Research Career Scientist Award, a VA Merit award to Dr. Alpini (5I01BX000574), a VA Merit Award (5I01BX002192) to Dr. Glaser, a VA Merit Award (1I01BX001724) to Dr. Meng from the United States (U.S.) Department of Veterans Affairs Biomedical Laboratory Research and Development Service, and the NIH grant DK076898 to Drs. Alpini, Meng and Glaser. This material is the result of work supported by resources at the Central Texas Veterans Health Care System. The views expressed in this article are those of the authors and do not necessarily represent the views of the Department of Veterans Affairs.
ASJC Scopus subject areas
- General