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Bulk RNA sequencing analysis of hepatocytes isolated from humanized liver

ABSTRACT: The goal of this experiment was to identify differentially expressed human genes in the hepatocyte fraction between two groups. Group A includes mice having only human hepatocytes and group B includes mice having both human hepatocytes and human Non-Parenchymal Cells (NPCs).

ORGANISM(S): Mus musculus

PROVIDER: GSE234757 | GEO | 2023/06/16

REPOSITORIES: GEO

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Single-cell RNA sequencing analysis of humanized liver

Project description:Single cell RNA sequencing of liver cells enriched for Non-Parenchymal cells (NPCs) to see the percentage of humanizaton, the gene signature and the heterogeneity of most types of human NPCs in a humanized liver.

2023-06-16 | GSE234758 | GEO

Adult hepatocytes direct liver organogenesis through non-parenchymal cell recruitment in the kidney

Project description:Background & Aims: Since the first account of the myth of Prometheus, the amazing regenerative capacity of the liver has fascinated researchers due to its enormous medical potential. Liver regeneration is promoted by multiple types of liver cells, including hepatocytes and liver non-parenchymal cells (NPCs), through the complex intercellular signaling. However, the liver organogenetic mechanism, especially the role of adult hepatocytes at ectopic sites, remains unknown. In this study, we demonstrate that hepatocytes alone spurred liver organogenesis to form an organ-sized complex 3D liver that exhibited native liver architecture and functions in the kidneys of mice. Methods: Isolated hepatocytes were transplanted under the kidney capsule of monocrotaline (MCT) and partial hepatectomy (PHx)-treated mice. To determine the origin of NPCs in neo-livers, hepatocytes were transplanted into MCT/PHx-treated green fluorescent protein (GFP) transgenic mice or wild-type mice transplanted with bone marrow (BM) cells isolated from GFP mice. Results: Hepatocytes engrafted at the subrenal space of mice underwent continuous growth in response to a chronic hepatic injury in the native liver. More than 1.5 yrs later, whole organ-sized liver tissues having a greater mass than those of the injured native liver had formed. Most remarkably, we revealed that at least three types of NPCs with similar phenotypic features to the liver NPCs were recruited from the host tissues including BM. The neo-livers in the kidney exhibited liver-specific functions and architectures, including sinusoidal vascular systems, zonal heterogeneity, and emergence of bile duct cells. Furthermore, the neo-livers successfully rescued the mice with lethal liver injury. Conclusion: Our data clearly showed that adult hepatocytes play a leading role as organizer cells in liver organogenesis at ectopic sites via NPC recruitment.

2017-12-25 | GSE99141 | GEO

AAV capsid prioritization in machine-perfused normal and steatotic human livers

Project description:Adeno-associated virus (AAV) vectors generated with five different capsids were intravenously injected into human livers undergoing normothermic machine perfusion (NMP). To assess the tropism of the five AAV vectors in the human livers, single-cell suspensions of hepatocytes and non-parenchymal cells (NPCs) were analyzed by single-cell RNA sequencing (scRNAseq). Differentially expressed genes were identified in AAV vector-transduced hepatocytes and unique cell populations.

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Phenotype and function of ex vivo expanded liver progenitor-like cells through biliary epithelial cells reversion

Project description:The current study combined genetic lineage tracing with in vitro small-molecule-based reprogramming to define liver progenitor cells (LPCs) derived from hepatic parenchymal and non-parenchymal cells (NPCs). tdTom+ hepatocytes were isolated from ROSA26tdTomato mice following AAV8-Tbg-Cre-mediated recombination, ALB/GFP-EpCAM- NPCs were isolated from AlbCreERT/R26GFP mice and EpCAM+ biliary epithelial cells (BECs) from wild-type intrahepatic bile ducts. A cocktail of small molecules was used to convert the isolated cells into LPCs.

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Gene expression of human hepatocytes isolated from chimeric mice with humanized liver

Project description:Chimeric mice with humanized livers are considered a useful animal model for predicting human drug metabolism and toxicity. In this study, the characteristics of fresh h-hepatocytes (cFHHs, PXB-cells®) isolated from chimeric mice (PXB-mice®) were evaluated in vitro to confirm their utility for drug development. The cFHHs cultured at high density (2.13 × 10^5 cells/cm2) displayed stable production of human albumin and cytochrome P450 (CYP) 3A activities for at least 21 days. The mRNA expression levels of 10 of 13 CYPs, UDP-glucuronosyltransferase (UGP), and transporters were maintained at >10% of the levels of freshly isolated cFHHs after 21 days. From 7-days cultured cFHHs at high density, many bile canaliculi were observed between cFHHs, and the accumulation of multidrug resistance-associated protein (MRP2) and bile salt export pump (BSEP) substrates in these bile canaliculi was clearly inhibited by cyclosporin A. We used microarrays to elucidate global gene expressions underlying this higher hepatic functions of high density cultured cFHHs (PXB-cells). Microarray analysis revealed that high density cultured cFHHs maintained high expressions of some transcription factors (HNF4α, PXR, and FXR) perhaps involved in the high CYP, UGT and transporter gene expressions of cFHHs.

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Gene expression of human hepatocytes isolated from chimeric mice with humanized liver

Project description:Gene expression of human hepatocytes isolated from chimeric mice with humanized liver

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Hepatocytes induces properties of wound healing type and regulatory macrophages in co-cultivated BMDM

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2023-07-07 | PXD012136 | Pride

Gene expression profile at single cell level of hepatocytes and non parenchymal cells (NPCs) isolated from a murine model of MASLD

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Bulk RNA sequencing analysis of hepatocytes isolated from humanized liver

Project description:Bulk RNA sequencing analysis of hepatocytes isolated from humanized liver

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Expression data of hepatocytes isolated from chimeric mouse livers repopulated with human hepatocytes and from normal human livers

Project description:We generated chimeric mice with livers that were predominantly repopulated with human hepatocytes. Hepatocytes were isolated from the chimeric mouse livers and their gene expressions were compared with hepatocytes isolated from normal human livers . Cluster and principal components analyses showed that gene expression profiles of hepatocytes from the chimeric mice and those from normal human livers were extremely closed. Additionally, we performed microarray experiments to examine gene expression in human tissues. This data was used for comparison with hepatocytes. A total of 22 tissues (bone marrow, cerebellum, colon, cortex, fetal brain, heart, kidney, liver, lung, pancreas, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea and uterus) were examined. The chimeric mice were generated by transplantation of 2 different donor hepatocytes. Hepatocytes were isolated from the mouse livers and normal human livers, and their cDNAs were used for microarray analysis. Total RNA isolated from human tissues and cell cultures were labeled and hybridized to the GeneChip Human Genome U133 Plus 2.0 Array according to the manufacturer's protocol.

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