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| Status |
Public on Aug 22, 2017 |
| Title |
Engraftment and Repopulation Potential of Late Gestation Fetal Rat Hepatocytes |
| Organism |
Rattus norvegicus |
| Experiment type |
Expression profiling by array
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| Summary |
Liver transplantation is the only therapeutic option for patients with end-stage liver disease. The shortage of donor organs has led to the search for alternative therapies to restore liver function and bridge patients to transplantation. Our previous work has shown that the proliferation of late gestation E19 fetal hepatocytes is mitogen-independent. This is manifested as differences in the control of ribosome biogenesis, global translation, cell cycle progression and gene expression. In the present study, we investigated whether E19 fetal hepatocytes would engraft and repopulate an injured adult liver.
Methods: Fetal hepatocytes were isolated using a monoclonal antibody against a hepatic surface protein, leucine amino peptidase (LAP). LAP+ and LAP- fractions were analyzed by immunofluorescence and microarray. Immunopurified E19 liver cells from DPPIV+ F344 rats were transplanted via splenic injection into partial hepatectomized DPPIV- rats that had been pretreated with mitomycin C.
Results: Phenotypic characterization of the LAP+ fetal hepatocytes revealed that more than a third of the isolated cells expressed ductal markers. Transcriptomic analysis revealed that these dual expressing cells represent a distinct subpopulation of less well differentiated hepatocytes. Transplanted immunopurified LAP+ late gestation fetal hepatocytes formed small hepatic, endothelial and occasional ductal colonies within one month. The average size of the colonies derived from the LAP+ cells increased so that by 10 months up to 35% of the liver was repopulated by donor-derived cells.
Conclusions: Our studies show that late gestation fetal hepatocytes, despite their being far along in the differentiation process, possess the capacity for extensive liver repopulation. This is likely related to the unexpected presence of a significant proportion of hepatocyte marker-positive cells maintaining a less well differentiated phenotype.
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| Overall design |
Two subpopulations of E19 fetal rat hepatocytes were isolated using monoclonal antibodies against the hepatic cell surface marker leucine aminopeptidase and the ducal marker OC.2. Adult hepatocytes were also isolated. RNA was isolated from triplicate biological replicates of fetal LAP+/OC.2- and LAP+/OC.2+ cells as well as adult hepatocytes using the mirvana kit. Affymetrix Rat ST 1.0 arrays were utilized.
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| Contributor(s) |
Boylan JM, Francois-Vaughan H, Gruppuso PA, Sanders JA |
| Citation(s) |
28749819 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 DK100301 |
The Fetal Hepatocyte Phenotype and Cell-Based Therapy for Liver Disease |
RHODE ISLAND HOSPITAL |
Jennifer A. Sanders |
| R01 HD024455 |
Novel Approaches to Understanding the Nutrient Regulation of Fetal Somatic Growth |
RHODE ISLAND HOSPITAL |
Jennifer A. Sanders |
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| Submission date |
Apr 17, 2017 |
| Last update date |
Nov 21, 2017 |
| Contact name |
Jennifer Ann Sanders |
| E-mail(s) |
Jennifer_Sanders@brown.edu
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| Organization name |
Rhode Island Hospital
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| Department |
Pediatric Endocrinology
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| Street address |
593 Eddy Street
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| City |
Providence |
| State/province |
RI |
| ZIP/Postal code |
02903 |
| Country |
USA |
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| Platforms (1) |
| GPL6247 |
[RaGene-1_0-st] Affymetrix Rat Gene 1.0 ST Array [transcript (gene) version] |
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| Samples (9)
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| GSM2579673 |
LAP+/OC.2+ E19 fetal hepatocytes bio rep 1 |
| GSM2579674 |
LAP+/OC.2+ E19 fetal hepatocytes bio rep 2 |
| GSM2579675 |
LAP+/OC.2+ E19 fetal hepatocytes bio rep 3 |
| GSM2579676 |
LAP+/OC.2- E19 fetal hepatocytes bio rep 1 |
| GSM2579677 |
LAP+/OC.2- E19 fetal hepatocytes bio rep 2 |
| GSM2579678 |
LAP+/OC.2- E19 fetal hepatocytes bio rep 3 |
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| Relations |
| BioProject |
PRJNA383149 |
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