scholarly journals Impact of Hepatic Stellate Cells in Scaffold-Free 3D-Bioprinting of the Liver Model

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Cutter J. Koehler ◽  
Wenjun Zhang ◽  
Julia R. Walsh ◽  
Raza A. Naqvi ◽  
Erika Gramelspacher ◽  
...  
Keyword(s):  
Hepatic Stellate Cells ◽  
Histological Analysis ◽  
Stellate Cells ◽  
3D Bioprinting ◽  
Mrna Transcription ◽  
Primary Hepatocytes ◽  
Albumin Secretion ◽  
Liver Sinusoidal Endothelial Cells ◽  
Liver Model ◽  
Organ Models

Background and Hypothesis: Hepatic stellate cells (HSC), which compromise ~15% of liver cells, are vital to hepatocellular function. Scaffold-free 3D-bioprinting (SF3DBP) offers an avenue for the creation of realistic organ models without the use of biomaterials. Therefore, we hypothesized that co-culturing primary hepatocytes with HSC in SF3DBP liver model would uphold hepatocyte function over time, providing us a better 3D-liver model for research.  Experimental Design: We used freshly thawed primary pig hepatocytes and immortalized pig HSC to generate spheroids with hepatocytes alone, HSC alone, or a combination of hepatocytes and HSC (2.5:1 ratio). Spheroids were formed using low adhesion plates, then characterized for distance from well center, diameter, roundness, and smoothness. A column of spheroids was printed using a Regenova 3D-bioprinter. Remaining loose spheroids are incubated over two weeks for albumin secretion, mRNA transcription, and histological analysis.  Results: Co-cultures of hepatocytes and HSC (2.5:1 ratio) formed spheroids within 48 hours, as did HSC only spheroids (Figure 1). Spheroids composed of only hepatocytes failed to form round spheroids. The combination spheroids increased in roundness and decreased in diameter between characterizations over 6 days.  Conclusion and Potential Impact: Spheroids proved too large to print at 48 hours but were successfully recognized and placed by the 3D-bioprinter. SF3DBP of combination spheroids would be viable by day 6. Optimization of spheroid composition using different cell ratios including HSC, hepatocytes, liver sinusoidal endothelial cells and fibroblasts, as well as optimization of spheroid incubation time will allow for production and printing of more advanced liver models. 

Reproducing human and cross-species drug toxicities using a Liver-Chip

2019 ◽  
Vol 11 (517) ◽  
pp. eaax5516 ◽  
Author(s):  
Kyung-Jin Jang ◽  
Monicah A. Otieno ◽  
Janey Ronxhi ◽  
Heng-Keang Lim ◽  
Lorna Ewart ◽  
...  
Keyword(s):  
Hepatic Stellate Cells ◽  
Animal Studies ◽  
Liver Toxicity ◽  
Stellate Cells ◽  
Hepatocellular Injury ◽  
Primary Hepatocytes ◽  
Liver Sinusoidal Endothelial Cells ◽  
Human Risk ◽  
Physiological Fluid ◽  
Species Specific

Nonclinical rodent and nonrodent toxicity models used to support clinical trials of candidate drugs may produce discordant results or fail to predict complications in humans, contributing to drug failures in the clinic. Here, we applied microengineered Organs-on-Chips technology to design a rat, dog, and human Liver-Chip containing species-specific primary hepatocytes interfaced with liver sinusoidal endothelial cells, with or without Kupffer cells and hepatic stellate cells, cultured under physiological fluid flow. The Liver-Chip detected diverse phenotypes of liver toxicity, including hepatocellular injury, steatosis, cholestasis, and fibrosis, and species-specific toxicities when treated with tool compounds. A multispecies Liver-Chip may provide a useful platform for prediction of liver toxicity and inform human relevance of liver toxicities detected in animal studies to better determine safety and human risk.

scholarly journals Platelet Interactions with Liver Sinusoidal Endothelial Cells and Hepatic Stellate Cells Lead to Hepatocyte Proliferation

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1243 ◽  
Author(s):  
Jeremy Meyer ◽  
Alexandre Balaphas ◽  
Pierre Fontana ◽  
Philippe Morel ◽  
Simon C. Robson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Liver Regeneration ◽  
Partial Hepatectomy ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Hepatocyte Proliferation ◽  
Liver Sinusoidal Endothelial Cells ◽  
Hepatocyte Growth

(1) Background: Platelets were postulated to constitute the trigger of liver regeneration. The aim of this study was to dissect the cellular interactions between the various liver cells involved in liver regeneration and to clarify the role of platelets. (2) Methods: Primary mouse liver sinusoidal endothelial cells (LSECs) were co-incubated with increasing numbers of resting platelets, activated platelets, or platelet releasates. Alterations in the secretion of growth factors were measured. The active fractions of platelet releasates were characterized and their effects on hepatocyte proliferation assessed. Finally, conditioned media of LSECs exposed to platelets were added to primary hepatic stellate cells (HSCs). Secretion of hepatocyte growth factor (HGF) and hepatocyte proliferation were measured. After partial hepatectomy in mice, platelet and liver sinusoidal endothelial cell (LSEC) interactions were analyzed in vivo by confocal microscopy, and interleukin-6 (IL-6) and HGF levels were determined. (3) Results: Co-incubation of increasing numbers of platelets with LSECs resulted in enhanced IL-6 secretion by LSECs. The effect was mediated by the platelet releasate, notably a thermolabile soluble factor with a molecular weight over 100 kDa. The conditioned medium of LSECs exposed to platelets did not increase proliferation of primary hepatocytes when compared to LSECs alone but stimulated hepatocyte growth factor (HGF) secretion by HSCs, which led to hepatocyte proliferation. Following partial hepatectomy, in vivo adhesion of platelets to LSECs was significantly increased when compared to sham-operated mice. Clopidogrel inhibited HGF secretion after partial hepatectomy. (4) Conclusion: Our findings indicate that platelets interact with LSECs after partial hepatectomy and activate them to release a large molecule of protein nature, which constitutes the initial trigger for liver regeneration.

scholarly journals Endothelin-1 in portal hypertension: The intricate role of hepatic stellate cells

2020 ◽  
Vol 245 (16) ◽  
pp. 1504-1512 ◽  
Author(s):  
Devaraj Ezhilarasan
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Portal Hypertension ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Chronic Liver ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Endothelin 1 ◽  
Activated Hepatic Stellate Cells

Portal hypertension is one of the most important cirrhosis-associated complications of chronic liver disease, leading to significant morbidity and mortality. After chronic liver injury, hepatic stellate cells reside in the perisinusoidal space activted and acquire a myofibroblast-like phenotype. The activated hepatic stellate cells act as both sources as well as the target for a potent vasoconstrictor endothelin-1. Activation of hepatic stellate cells plays a vital role in the onset of cirrhosis by way of increased extracellular matrix production and the enhanced contractile response to vasoactive mediators such as endothelin-1. In fibrotic/cirrhotic liver, activated hepatic stellate cells produce endothelin-1 leading to an imbalance between pro and antifibrotic factors responsible for enormous extracellular matrix synthesis. Thus, extracellular matrix deposition in the perisinusoidal space further augments liver stiffness and elevates the vascular tone and portal hypertension. Portal hypertension is a complex process modulated by several cell types like hepatic stellate cells, liver sinusoidal endothelial cells, Kupffer cells, injured hepatocytes, immune cells, and biliary epithelial cells. Therefore, targeting a single cell type may not be useful for regression of cirrhosis and portal hypertension. Nevertheless, numerous findings indicate that functionally liver sinusoidal endothelial cells and hepatic stellate cells closely regulate the sinusoidal blood flow via synthesis of several vasoactive molecules including endothelin-1, and hence targeting these cells with novel pharmacological agents may offer promising results. Impact statement Portal hypertension is pathologically defined as increase of portal venous pressure, mainly due to chronic liver diseases such as fibrosis and cirrhosis. In fibrotic liver, activated hepatic stellate cells increase their contraction in response to endothelin-1 (ET-1) via autocrine and paracrine stimulation from liver sinusoidal endothelial cells and injured hepatocytes. Clinical studies are limited with ET receptor antagonists in cirrhotic patients with portal hypertension. Hence, studies are needed to find molecules that block ET-1 synthesis. Accumulation of extracellular matrix proteins in the perisinusoidal space, tissue contraction, and alteration in blood flow are prominent during portal hypertension. Therefore, novel matrix modulators should be tested experimentally as well as in clinical studies. Specifically, tumor necrosis factor-α, transforming growth factor-β1, Wnt, Notch, rho-associated protein kinase 1 signaling antagonists, and peroxisome proliferator-activated receptor α and γ, interferon-γ and sirtuin 1 agonists should be tested elaborately against cirrhosis patients with portal hypertension.

scholarly journals Targeting integrin αvβ3 by a rationally designed protein for chronic liver disease treatment

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Ravi Chakra Turaga ◽  
Ganesh Satyanarayana ◽  
Malvika Sharma ◽  
Jenny J. Yang ◽  
Shiyuan Wang ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Hepatic Stellate Cells ◽  
Flow Resistance ◽  
Integrin Αvβ3 ◽  
Stellate Cells ◽  
Chronic Liver ◽  
Chronic Liver Diseases ◽  
Liver Sinusoidal Endothelial Cells ◽  
Disease Treatment ◽  
Abnormal Accumulation

AbstractChronic Liver Diseases (CLD) are characterized by abnormal accumulation of collagen fibrils, neo-angiogenesis, and sinusoidal remodeling. Collagen deposition along with intrahepatic angiogenesis and sinusoidal remodeling alters sinusoid structure resulting in portal hypertension, liver failure, and other complications. Efforts were made to develop treatments for CLDs. However, the success of such treatments is limited and unpredictable. We report a strategy for CLD treatment by induction of integrin αvβ3 mediated cell apoptosis using a rationally designed protein (ProAgio). ProAgio is designed to target integrin αvβ3 at a novel site. Integrin αvβ3 is highly expressed in activated Hepatic Stellate Cells (HSC), angiogenic endothelium, and capillarized Liver Sinusoidal Endothelial Cells (LSEC). ProAgio induces apoptosis of these disease causative cells. Tests with liver fibrosis mouse models demonstrate that ProAgio reverses liver fibrosis and relieves blood flow resistance by depleting activated HSC and capillarized LSEC. Our studies demonstrate an effective approach for CLD treatment.

scholarly journals Nano delivery of simvastatin targets liver sinusoidal endothelial cells to remodel tumor microenvironment for hepatocellular carcinoma

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhuo Yu ◽  
Jianfeng Guo ◽  
Yun Liu ◽  
Menglin Wang ◽  
Zhengsheng Liu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Endothelial Cells ◽  
Tumor Microenvironment ◽  
Hepatic Stellate Cells ◽  
Targeted Delivery ◽  
Stellate Cells ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Fibrotic Liver ◽  
Stromal Microenvironment

Abstract Background Hepatocellular carcinoma (HCC) developed in fibrotic liver does not respond well to immunotherapy, mainly due to the stromal microenvironment and the fibrosis-related immunosuppressive factors. The characteristic of liver sinusoidal endothelial cells (LSECs) in contributing to fibrosis and orchestrating immune response is responsible for the refractory to targeted therapy or immunotherapy of HCC. We aim to seek a new strategy for HCC treatment based on an old drug simvastatin which shows protecting effect on LSEC. Method The features of LSECs in mouse fibrotic HCC model and human HCC patients were identified by immunofluorescence and scanning electron microscopy. The effect of simvastatin on LSECs and hepatic stellate cells (HSCs) was examined by immunoblotting, quantitative RT-PCR and RNA-seq. LSEC-targeted delivery of simvastatin was designed using nanotechnology. The anti-HCC effect and toxicity of the nano-drug was evaluated in both intra-hepatic and hemi-splenic inoculated mouse fibrotic HCC model. Results LSEC capillarization is associated with fibrotic HCC progression and poor survival in both murine HCC model and HCC patients. We further found simvastatin restores the quiescence of activated hepatic stellate cells (aHSCs) via stimulation of KLF2-NO signaling in LSECs, and up-regulates the expression of CXCL16 in LSECs. In intrahepatic inoculated fibrotic HCC mouse model, LSEC-targeted nano-delivery of simvastatin not only alleviates LSEC capillarization to regress the stromal microenvironment, but also recruits natural killer T (NKT) cells through CXCL16 to suppress tumor progression. Together with anti-programmed death-1-ligand-1 (anti-PD-L1) antibody, targeted-delivery of simvastatin achieves an improved therapeutic effect in hemi-splenic inoculated advanced-stage HCC model. Conclusions These findings reveal an immune-based therapeutic mechanism of simvastatin for remodeling immunosuppressive tumor microenvironment, therefore providing a novel strategy in treating HCC. Graphical Abstract

scholarly journals Hepatocyte CREBZF-osteopontin axis controls fibrosis in nonalcoholic steatohepatitis

2021 ◽  
Author(s):  
Fengguang Ma ◽  
Yuxiao Liu ◽  
Zhimin Hu ◽  
Yaqian Xue ◽  
Zhengshuai Liu ◽  
...  
Keyword(s):  
Nonalcoholic Steatohepatitis ◽  
Hepatic Stellate Cells ◽  
Matrix Protein ◽  
Stellate Cells ◽  
Extracellular Matrix Protein ◽  
Chow Diet ◽  
Primary Hepatocytes ◽  
Adeno Associated Virus

Abstract Nonalcoholic steatohepatitis (NASH) has emerged as a leading cause of chronic liver disease. The incomplete understanding of NASH fibrosis limits pharmacotherapy development. Here we report a molecular link between hepatocytes and hepatic stellate cells (HSCs) in regulating the progression of liver fibrosis via CREBZF-osteopontin (OPN) axis. Hepatocyte-specific CREBZF knockout (CREBZF LKO) mice and their wild-type littermates were divided into groups that were placed on AMLN, MCD or chow diet. Mouse primary hepatocytes were treated with 250 μM PA and 10 ng/ml TNFα, the conditioned medium was collected and then transferred to HSC-T6 cells for 24 hours. Adeno-associated virus-mediated overexpression of OPN or CREBZF was performed in mice via tail vail injection. Human studies have shown that deregulation of CREBZF is associated with pathogenesis of hepatic steatosis and dyslipidemia. Here, we show that CREBZF is markedly elevated in livers of NASH mice, whereas hepatocyte-specific CREBZF knockout mice are prevented from AMLN or MCD diet-induced hepatic inflammation, liver injury and fibrosis. In vivo and in vitro mechanistic studies revealed that a key mechanism linking hepatocyte CREBZF to NASH fibrosis is miR-6964-3p-mediated inhibition of OPN, an extracellular matrix protein that activates fibrogenic genes in hepatic stellate cells. Moreover, the reduction of NASH phenotypes in CREBZF LKO mice was reversed by adeno-associated virus-mediated overexpression of OPN. Thus, CREBZF-OPN axis represents a previously unrecognized intrahepatic crosstalk in the liver that triggers fibrosis progression and contributes to the severity of NASH.

scholarly journals Hepatic Stellate Cells Improve Engraftment of Human Primary Hepatocytes: A Preclinical Transplantation Study in an Animal Model

2015 ◽  
Vol 24 (12) ◽  
pp. 2557-2571 ◽  
Author(s):  
Ange-Clarisse Dusabineza ◽  
Mustapha Najimi ◽  
Noémi Van Hul ◽  
Vanessa Legry ◽  
Dung Ngoc Khuu ◽  
...  
Keyword(s):  
Animal Model ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Primary Hepatocytes ◽  
Human Primary Hepatocytes ◽  
Transplantation Study

scholarly journals Role of Liver-Mediated Tolerance in Nanoparticle-Based Tumor Therapy

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1985
Author(s):  
Maximiliano L. Cacicedo ◽  
Carolina Medina-Montano ◽  
Leonard Kaps ◽  
Cinja Kappel ◽  
Stephan Gehring ◽  
...  
Keyword(s):  
Hepatic Stellate Cells ◽  
Liver Diseases ◽  
Potential Role ◽  
Current Knowledge ◽  
Tumor Therapy ◽  
Stellate Cells ◽  
Liver Sinusoidal Endothelial Cells ◽  
Parenchymal Cells ◽  
Systemic Application

In the last decades, the use of nanocarriers for immunotherapeutic purposes has gained a lot of attention, especially in the field of tumor therapy. However, most types of nanocarriers accumulate strongly in the liver after systemic application. Due to the default tolerance-promoting role of liver non-parenchymal cells (NPCs), Kupffer cells (KCs), liver sinusoidal endothelial cells (LSECs), and hepatic stellate cells (HSCs), their potential role on the immunological outcome of systemic nano-vaccination approaches for therapy of tumors in the liver and in other organs needs to be considered. Concerning immunological functions, KCs have been the focus until now, but recent studies have elucidated an important role of LSECs and HSCs as well. Therefore, this review aims to summarize current knowledge on the employment of nanocarriers for immunotherapeutic therapy of liver diseases and the overall role of liver NPCs in the context of nano-vaccination approaches. With regard to the latter, we discuss strategies on how to address liver NPCs, aiming to exploit and modulate their immunological properties, and alternatively how to avoid unwanted engagement of nano-vaccines by liver NPCs for tumor therapy.

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