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Hepatocyte Isolation

Tong Cao, Jing Pan, Xiulian Li, Yanli He, Yifei Jiang, Yajing Chang, Qi Zhang, Ying Lan, Hua Wang, Wenjie Jiao, Zibin Tian, Lijuan Zhang
BACKGROUND/AIMS: The up-regulation of hepatocyte growth factor/receptor, HGF/Met, signal transduction is observed in most of human cancers. Specific heparan sulfate structures enhance the HGF/Met signaling at both cell and animal-based model systems. Biochemical studies indicate that heparan sulfate interacts with HGF and a natural occurring splicing variant NK1 of HGF with similar affinity. However, it is currently unknown if cell surface heparan sulfate binds to Met at physiological conditions and if specific cell surface heparan sulfate structures are required for effective HGF/Met or NK1/Met signaling...
August 14, 2018: Cellular Physiology and Biochemistry
Jianfeng Tang, Xiaoming Wang, Kezhe Tan, Hongtao Zhu, Youming Zhang, Weili Ouyang, Xueqing Liu, Zhaoping Ding
BACKGROUND: Injury may induce a sequential activation of intrinsic reparative activity that supports the maintenance of tissue homeostasis. METHOD: In the present experiments, we investigated whether myocardial infarction (MI) was able to reinstate the expression of Wilms' tumor factor 1 (WT1) as a key hallmark of fetal reprograming in the pericardial adipose-derived stem cells (pADSC). We characterized the immunophenotypical markers, cardiac potential, and reparative activity of WT1-expressing pADSC (WT1pos ) isolated MI Wistar rats with an intact pericardial sac in which cardiac transudate was accumulated, sampled, and analyzed...
August 13, 2018: Stem Cell Research & Therapy
Caiyun Ma, Yu Guo, Hebao Wen, Yanjie Zheng, Leiqi Tan, Xiangchen Li, Chunjing Wang, Weijun Guan, Changqing Liu
Adipose tissue-derived mesenchymal stem cells (ADSCs) play a crucial role in the field of regenerative medicine and tissue repair for its own unique features. However, up to date, the isolation and characterizations of multidifferentiation potentials of goose ADSCs are still uncertain. In this study, we successfully isolated ADSCs from goose inguinal groove in vitro for the first time and also attempted to unravel its fundamental differentiation potentials and genetic characteristics. The results showed that isolated ADSCs exhibited a typical fibroblast-like morphology and high proliferative potential, could be passaged for at least 40 passages and maintained high hereditary stability with more than 92...
August 13, 2018: DNA and Cell Biology
Xiliang Du, Taiyu Shen, Heyuan Wang, Xia Qin, Dongmei Xing, Qianqian Ye, Zhen Shi, Zhiyuan Fang, Yiwei Zhu, Yuchen Yang, Zhicheng Peng, Chenxu Zhao, Bin Lv, Xiaobing Li, Guowen Liu, Xinwei Li
The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. Fatty liver is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and mitochondria in dairy cows with mild fatty liver, liver and blood samples were collected from healthy cows (n = 15) and cows with mild fatty liver (n = 15). To determine the effects of palmitic acids (PA), one of the major component of fatty acids, on lipid metabolism and mitochondria in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM)...
August 9, 2018: Journal of Dairy Science
Jian Wang, Song Wang, Chongjia Yan, Yunxia Deng, Zhiwei Huang, Ping Shi
As a lipin family founding member, lipin1 exerts dual functions as a phosphatidate phosphatase enzyme and/or a co-transcriptional regulator in lipid metabolism. In fact, it is also involved in many other cell processes. In this study, we utilized pull down assay coupled with mass spectrometry (MS) to unravel protein-protein interaction networks of lipin1 in 293T human embryonic kidney cells. Pull-down assay on the Ni2+ -chelating column was used to isolate lipin1 complexes from 293T cells transfected with 6-His tagged lipin1...
August 9, 2018: IUBMB Life
Charles E McCall, Manal Zabalawi, Tiefu Liu, Ayana Martin, David L Long, Nancy L Buechler, Rob J W Arts, Mihai Netea, Barbara K Yoza, Peter W Stacpoole, Vidula Vachharajani
Limited understanding of the mechanisms responsible for life-threatening organ and immune failure hampers scientists' ability to design sepsis treatments. Pyruvate dehydrogenase kinase 1 (PDK1) is persistently expressed in immune-tolerant monocytes of septic mice and humans and deactivates mitochondrial pyruvate dehydrogenase complex (PDC), the gate-keeping enzyme for glucose oxidation. Here, we show that targeting PDK with its prototypic inhibitor dichloroacetate (DCA) reactivates PDC; increases mitochondrial oxidative bioenergetics in isolated hepatocytes and splenocytes; promotes vascular, immune, and organ homeostasis; accelerates bacterial clearance; and increases survival...
August 9, 2018: JCI Insight
Haeyoung Zhang, Abdul Basit, Diana Busch, King Yabut, Deepak Kumar Bhatt, Marek Drozdzik, Marek Ostrowski, Albert Li, Carol Collins, Stefan Oswald, Bhagwat Prasad
Protein abundance and activity of UGT2B17, a highly variable drug- and androgen-metabolizing enzyme, was quantified in microsomes, S9, and primary cells isolated from human liver and intestine by a validated LC-MS/MS methods. UGT2B17 protein abundance showed >160-fold variation (mean ± SD, 1.7 ± 2.7 pmol/mg microsomal protein) in adult human liver microsomes (n=26) and significant correlation (r2 = 0.77, p<0.001) with TG formation. Primary role of UGT2B17 in TG formation compared to UGT2B15 was confirmed by performing the activity assay in UGT2B17 gene deletion samples and with a selective UGT2B17 inhibitor, imatinib...
August 4, 2018: Biochemical Pharmacology
Ryuji Watari, Motoharu Kakiki, Ayumi Oshikata, Toshiaki Takezawa, Chihiro Yamasaki, Yuji Ishida, Chise Tateno, Yukie Kuroda, Seiichi Ishida, Kazutomi Kusano
During drug discovery, in vitro models are used to predict the in vivo pharmacokinetic and toxicological properties of drug candidates in humans. However, the conventional method of culturing human hepatocytes as monolayers does not necessarily replicate biologic reactions and does not support liver-specific functions, such as cytochrome P450 (CYP) activities, for prolonged periods. To remedy these problems and thus increase and prolong hepatic functions, we developed a culture system comprising a collagen vitrigel membrane (CVM) chamber and PXB-cells®, fresh hepatocytes isolated from liver-humanized chimeric mice (PXB-mice®)...
2018: Journal of Toxicological Sciences
Ling Yang, Weijun Wang, Xiaozhan Wang, Jinfang Zhao, Li Xiao, Wenfang Gui, Huiqian Fan, Jing Xia, Zhonglin Li, Jingjing Yan, Afnan Alasbahi, Qingjing Zhu, Xiaohua Hou
Hepatic ischemia/reperfusion (I/R) is a major challenge for liver surgery and specific severe conditions of chronic liver disease. Current surgical and pharmacological strategies are limited to improve liver function after hepatic I/R injury. Thus, an in-depth understanding of the liver I/R mechanism is pivotal to develop new therapeutic methods. The cellular repressor of E1A-stimulated genes (CREG), a key regulator of cellular proliferation, exerts protective roles in cardiovascular diseases and participates in lipid accumulation and inflammatory response in the liver...
August 4, 2018: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
Ruy A Louzada, Denise P Carvalho
Thyroxine (T4) and 3,5,3'-triiodothyronine (T3) are secreted by the thyroid gland, while T3 is also generated from the peripheral metabolism of T4 by iodothyronine deiodinases types I and II. Several conditions like stress, diseases, and physical exercise can promote changes in local TH metabolism, leading to different target tissue effects that depend on the presence of tissue-specific enzymatic activities. The newly discovered physiological and pharmacological actions of T4 and T3 metabolites, such as 3,5-diiodothyronine (3,5-T2), and 3-iodothyronamine (T1AM) are of great interest...
2018: Frontiers in Endocrinology
Liana Monteiro da Fonseca Cardoso, Lucio Filgueiras Pacheco Moreira, Marcelo Alves Pinto, Andrea Henriques-Pons, Luiz Anastácio Alves
Background and Aims: Acute liver failure (ALF) is a severe syndrome with an elevated mortality rate, ranging from 40 to 80 %. Currently, liver transplantation is the only definitive treatment for these patients and new therapies aiming to treat ALF include artificial organs implant and stem cells therapy, for example. However, a major limitation of liver donors exists. Living donor liver transplantation (LDLT), split liver transplantation (SLT), and domino liver transplantation (DLT) are some of the available alternatives to treat ALF patients, but these do not reduce the number of patients on waiting lists...
2018: Canadian Journal of Gastroenterology & Hepatology
Alan Moreira de Araujo, Maísa Mota Antunes, Matheus Silvério Mattos, Ariane Barros Diniz, Débora Moreira Alvarenga, Brenda Naemi Nakagaki, Érika de Carvalho, Viviane Aparecida Souza Lacerda, Raquel Carvalho-Gontijo, Jorge Goulart, Kassiana Mafra, Maria Alice Freitas-Lopes, Hortência Maciel de Castro Oliveira, Camila Miranda Dutra, Bruna Araújo David, Aristóbolo Mendes Silva, Valerie Quesniaux, Bernhard Ryffel, Sergio Costa Oliveira, Glen N Barber, Daniel Santos Mansur, Thiago Mattar Cunha, Rafael Machado Rezende, André Gustavo Oliveira, Gustavo Batista Menezes
Hepatocytes may rupture after a drug overdose, and their intracellular contents act as damage-associated molecular patterns (DAMPs) that lead to additional leukocyte infiltration, amplifying the original injury. Necrosis-derived DNA can be recognized as a DAMP, activating liver non-parenchymal cells (NPCs). We hypothesized that NPCs react to DNA by releasing interferon (IFN)-1, which amplifies acetaminophen (APAP)-triggered liver necrosis. We orally overdosed different knockout mouse strains to investigate the pathways involved in DNA-mediated amplification of APAP-induced necrosis...
July 27, 2018: Cells
Yasushi Miura, Satoshi Matsui, Naoko Miyata, Kenichi Harada, Yamato Kikkawa, Masaki Ohmuraya, Kimi Araki, Shinya Tsurusaki, Hitoshi Okochi, Nobuhito Goda, Atsushi Miyajima, Minoru Tanaka
Under chronic or severe liver injury, liver progenitor cells (LPCs) of biliary origin are known to expand and contribute to the regeneration of hepatocytes and cholangiocytes. This regeneration process is called ductular reaction (DR), which is accompanied by dynamic remodeling of biliary tissue. Although the DR shows apparently distinct mode of biliary extension depending on the type of liver injury, the key regulatory mechanism remains poorly understood. Here, we show that Lutheran (Lu)/Basal cell adhesion molecule (BCAM) regulates the morphogenesis of DR depending on liver disease models...
July 30, 2018: ELife
Manal M Kamel, Hanan G El Baz, Zeinab Demerdash, Salwa Hassan, Faten Salah, Wafaa A Mansour, Olfat Hammam, Shimaa Atta, Ali Bayoumi, Marwa Hassan, Soheir Mahmoud
BACKGROUND: The liver is one of the major target organs for which cell-based therapies are very promising. The limitations of various cellular therapies, including bone marrow (BM)-derived mesenchymal stem cells (MSCs), urges the exploration of stem cell sources more suitable for transplantation. Human umbilical cord blood (HUCB) can overcome these drawbacks with a favorable reparative outcome. OBJECTIVES: The aim of this study was to evaluate the therapeutic potential of MSCs in 2 groups of chronic liver injury experimental models...
July 26, 2018: Advances in Clinical and Experimental Medicine: Official Organ Wroclaw Medical University
Shinichiro Minami, Kouichi Miura, Mitsuaki Ishioka, Naoki Morimoto, Norio Isoda, Hironori Yamamoto, Katsunori Iijima
AIM: High concentrations of homocysteine are believed to induce lipid synthesis and cell injury through endoplasmic reticulum (ER) stress in metabolic syndrome. However, homocysteine may be used to improve steatohepatitis induced by choline deficiency, in which methyl donors are decreased. The aim of the present study was to clarify the role of the physiological concentration of homocysteine in the development of steatohepatitis induced by choline deficiency. METHODS: Wild-type (WT) mice were fed a choline-deficient amino acid-defined (CDAA) diet with or without homocysteine supplementation for 24 weeks...
July 26, 2018: Hepatology Research: the Official Journal of the Japan Society of Hepatology
Sten Orrenius
My research activity started with studies on drug metabolism in rat liver microsomes in the early 1960s. The CO-binding pigment (cytochrome P450) had been discovered a few years earlier and was subsequently found to be involved in steroid hydroxylation in adrenal cortex microsomes. Our early studies suggested that it also participated in the oxidative demethylation of drugs catalyzed by liver microsomes, and that prior treatment of the animals with phenobarbital caused increased levels of the hemoprotein in the liver, and similarly enhanced rates of drug metabolism...
July 25, 2018: Annual Review of Pharmacology and Toxicology
Daniel Todt, Nora Moeller, Dimas Praditya, Volker Kinast, Martina Friesland, Michael Engelmann, Lieven Verhoye, Ibrahim M Sayed, Patrick Behrendt, Viet Loan Dao Thi, Philip Meuleman, Eike Steinmann
Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and a member of the genus Orthohepevirus in the family Hepeviridae. HEV infections are the common cause of acute hepatitis but can also take chronic courses. Ribavirin is the treatment of choice for most patients and type I interferon (IFN) has been evaluated in a few infected transplantation patients in vivo. However, no effective and specific treatments against HEV infections are currently available. In this study, we evaluated the natural compound silvestrol, isolated from the plant Aglaia foveolata, and known for its specific inhibition of the DEAD-box RNA helicase eIF4A in state-of-the-art HEV experimental model systems...
September 2018: Antiviral Research
Hengli Zhao, Qiuju Han, Nan Lu, Dongqing Xu, Zhigang Tian, Jian Zhang
The HMBOX1 (Homeobox Containing 1) gene was first isolated from the human pancreatic cDNA libraries and is widely expressed in many tissues. Previously, we detected high expression of HMBOX1 in the liver, but its function was unclear. In this study, hepatocyte-specific HMBOX1 knockout mice (Hm△hep mice) were generated and used to characterize the function of HMBOX1 in the LPS/D-GalN-induced acute liver failure model. HMBOX1-knockout exhibits exacerbated liver injury induced by LPS/D-GalN, accompanied with high levels of inflammatory cytokines both in the liver and in circulation...
July 19, 2018: Molecular Immunology
Chenxuan Liu, Guangwei Xu, Zhenchao Gao, Zhongmin Zhou, Guilan Guo, Dan Li, Zhiyi Jing, Jianhua Sui, Wenhui Li
Sodium taurocholate cotransporting polypeptide (NTCP) is a functional receptor for human hepatitis B virus (HBV) and its satellite virus Hepatitis D virus (HDV). Physiologically, NTCP is responsible for the majority of sodium-dependent bile acids uptake by hepatocytes. The p.Ser267Phe (S267F) variant of NTCP is a single nucleotide polymorphism (SNP) previously found to cause substantial loss of ability to support HBV and HDV infection and its taurocholic acid uptake function in vitro. Intriguingly, ten individuals were identified as S267F homozygotes in population studies of chronic hepatitis B (CHB) patients...
September 2018: Virology
Franziska Paech, Vanessa F Abegg, Urs Duthaler, Luigi Terracciano, Jamal Bouitbir, Stephan Krähenbühl
Reports concerning hepatic mitochondrial toxicity of sunitinib are conflicting. We therefore decided to conduct a toxicological study in mice. After having determined the highest dose that did not affect nutrient ingestion and body weight, we treated mice orally with sunitinib (7.5 mg/kg/day) for 2 weeks. At the end of treatment, peak sunitinib plasma concentrations were comparable to those achieved in humans and liver concentrations were approximately 25-fold higher than in plasma. Sunitinib did not affect body weight, but increased plasma ALT activity 6-fold...
July 18, 2018: Toxicology
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