Abstract:
INTRODUCTION:Primary porcine hepatocytes are commonly, used in bioartificial liver devices and for in vitro studies of hepatocyte function. Although in vivo isolation of porcine hepatocytes can give high yield and viability, such methods are time-consuming and expensive, requiring specialist surgical facilities. AIM:To develop a simple, low-cost, high viability, high yield, reproducible ex vivo method for obtaining functional porcine hepatocytes for use in bioartificial liver systems. METHODS:Weanling piglets (12 kg) were killed with pentobarbitone sodium, the infra-hepatic inferior vena cava was clamped and the supra-hepatic inferior vena cava cannulated. The whole liver was retrogradely perfused in situ with cold saline and excised, followed by an ex vivo open-loop and re-circulating perfusion method (at 37 degrees C) in five steps. The liver was disrupted, sequentially filtered in washing buffer, purified by centrifugation and resuspended in Williams E medium. Viability and cell number were assessed using trypan blue exclusion. The cells were subsequently cultured in serum-free chemically-defined medium and function was assessed. RESULTS:The time interval from when the animals were killed to the final cell wash was 105+/-5 min (n = 20). Cell viability was 85+/-6% with a yield of (2.4+/-0.5) x 10(10) from 12+/-1 kg piglets using 0.03% (w/v) collagenase (n = 20). Hepatocytes from all isolations were successfully plated and grown in monolayer culture. In freshly isolated hepatocytes (day 0) total protein content (TP) was 1.2+/-0.1 mg/10(6) cells (n = 5) and 1.2+/-0.3 mg/10(6) cells (n = 5) for day 2 monolayer cultures, corresponding to approximately 9x10(6) hepatocytes per dish. The percentage of total LDH released into the medium was 13+/-4% for day 0 and 8+/-4% at day 2; conversely, intracellular LDH activities were 87+/-4% and 92+/-4% of the total, respectively. The urea synthesis rate was 196+/-36 nmol/h/mg total protein at day 0 (n = 5) and 292+/-62 nmol/h/mg protein (n = 9) at day 2. The total P450 content was 99+/-11 pmol/mg total protein for fresh cells (n = 5) and maintained at 89+/-35 pmol/mg total protein in day 2 cultures. CONCLUSIONS:This ex vivo method provides a high viability, high yield, cost-effective and rapid technique for isolating functional porcine hepatocytes with high plating efficiency, which compares favourably with results obtained using complex in vivo techniques.
journal_name
Eur J Gastroenterol Hepatoljournal_title
European journal of gastroenterology & hepatologyauthors
Nelson LJ,Newsome PN,Howie AF,Hadoke PW,Dabos KJ,Walker SW,Hayes PC,Plevris JNdoi
10.1097/00042737-200012080-00012subject
Has Abstractpub_date
2000-08-01 00:00:00pages
923-30issue
8eissn
0954-691Xissn
1473-5687journal_volume
12pub_type
杂志文章abstract:BACKGROUND AND AIMS:Current guidelines for chronic hepatitis B (CHB) patients are to undergo surveillance for hepatocellular carcinoma (HCC) with 6-month ultrasonography. We aimed to compare detection rates of very-early-stage HCC in two groups: group A, undergoing 6-month ultrasonography versus group B, undergoing 6-m...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/MEG.0000000000001870
更新日期:2020-08-14 00:00:00
abstract:OBJECTIVE:To examine the impact of ileorectal anastomosis on vitamin B12 absorption, as measured by the Schilling test, in patients with Crohn's disease. PATIENTS:Eighty-two patients with Crohn's disease who had undergone ileorectal anastomosis. METHODS:Of the 82 patients with Crohn's disease and ileorectal anastomos...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:
更新日期:1995-05-01 00:00:00
abstract:OBJECTIVES:Hepatitis C virus (HCV) is the major cause of cryoglobulinemia. Direct-acting antivirals (DAAs) have markedly changed the therapeutic outcomes in the treatment of patients with HCV. We evaluate the efficacy, safety, immunological, and clinical response of different DAA regimens in HCV-cryoglobulinemia. PATI...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/MEG.0000000000001239
更新日期:2018-10-01 00:00:00
abstract:OBJECTIVE:Celiac disease (CD) is a lifelong, chronic, immune-mediated, inflammatory small bowel disorder, precipitated by exposure to dietary gluten and related proteins in genetically predisposed individuals. Recent studies have shed new light on the importance of inflammation in the pathogenesis of arterial stiffness...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/MEG.0000000000000437
更新日期:2015-10-01 00:00:00
abstract:OBJECTIVES:To investigate the effect of membrane lipid composition on the susceptibility to bile salt damage and protection. DESIGN:Artificial model cholesterol/phospholipid (c/p) membranes (vesicles) with a varying cholesterol (0-15 mM) and phospholipid content (3-30 mM), and with a c/p ratio ranging up to 1.70, were...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/00042737-199705000-00012
更新日期:1997-05-01 00:00:00
abstract::Brucellosis is a zoonosis with a worldwide distribution, and still endemic in Greece. Although hepatic involvement is common, cholecystitis and hepatic abscesses represent rare complications of the disease. Hematological manifestations of brucellosis include various abnormalities such as anemia and leucopenia, but the...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/MEG.0b013e3282ef9493
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abstract:BACKGROUND:A 7 day treatment scheme based on rabeprazole/levofloxacin/amoxycillin or tinidazole achieved an eradication rate over 90%. However, the combination of drugs and duration of treatment for the correct use of levofloxacin in the eradication of are still unclear. OBJECTIVE:To compare the efficacy and tolerabil...
journal_title:European journal of gastroenterology & hepatology
pub_type: 临床试验,杂志文章,随机对照试验
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abstract:BACKGROUND AND AIMS:Traditional definitions of healthcare utilization (HCU) emphasize clinical visits and procedures. Clinic calls, an understudied form of HCU, occur with high frequency. Understanding and examining predictors of HCU, such as disease activity and parent distress, may help reduce overutilization. METHO...
journal_title:European journal of gastroenterology & hepatology
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abstract::Prophylaxis with norfloxacin has been shown to be effective in preventing spontaneous bacterial peritonitis (SBP) in liver cirrhosis. This therapy is associated with the appearance of quinolone-resistant Escherichia coli. However, only four cases of SBP caused by quinolone-resistant E. coli have been reported so far. ...
journal_title:European journal of gastroenterology & hepatology
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doi:10.1097/00042737-200201000-00014
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journal_title:European journal of gastroenterology & hepatology
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abstract:BACKGROUND:Hepatitis C virus (HCV) is a major cause of chronic hepatitis and hepatic fibrosis. Liver biopsy, because of its limitations and risks, might be considered an imperfect gold standard for assessing the severity of chronic liver diseases. In this study, we aimed to prospectively validate FibroTest (FT) and Act...
journal_title:European journal of gastroenterology & hepatology
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abstract:BACKGROUND:Liver steatosis (LS) and chronic infection with hepatitis B virus (HBV) are two common causes of chronic liver disease in Iran. LS is also a common histopathological feature of chronic hepatitis B (CHB). Association of chronic HBV infection and LS has not been extensively studied. AIM:We investigated the pr...
journal_title:European journal of gastroenterology & hepatology
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abstract:BACKGROUND:We recently reported that (Lamiaceae) may alleviate CCl(4)-induced acute hepatotoxicity in rats, possibly blocking the formation of free radicals generated during CCl(4) metabolism. Carnosol, one of the main constituents of Rosmarinus, has been shown to have antioxidant and scavenging activities. Therefore, ...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/00042737-200209000-00011
更新日期:2002-09-01 00:00:00
abstract::A 38-year-old woman presented with a mucosal gastric carcinoma measuring 0.7 x 0.5 cm and para-aortic lymph node metastasis. Radiographic and endoscopic studies showed a small depressed lesion on the anterior border of the gastric angle, which was classified as a type II c + III lesion. Histological examination of the...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章,评审
doi:
更新日期:1999-03-01 00:00:00
abstract:OBJECTIVE:To determine the efficacy of lansoprazole 30 mg given in the morning compared with high-dose ranitidine 300 mg twice daily in the treatment of patients with oesophageal strictures. DESIGN:A multicentre, outpatient, double-blind, parallel group, prospectively randomized clinical trial. PATIENTS:One hundred a...
journal_title:European journal of gastroenterology & hepatology
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doi:
更新日期:1996-05-01 00:00:00
abstract:AIM:Although liver biopsy has long been considered the gold standard for staging fibrosis, because of the disadvantages and risks of biopsy, several noninvasive processes such as serum biomarkers have been introduced for the assessment of liver fibrosis. The aim of this study was to assess the diagnostic value of serum...
journal_title:European journal of gastroenterology & hepatology
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journal_title:European journal of gastroenterology & hepatology
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doi:10.1097/MEG.0b013e3282f1623d
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journal_title:European journal of gastroenterology & hepatology
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journal_title:European journal of gastroenterology & hepatology
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journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/00042737-200105000-00009
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journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章,meta分析,评审
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更新日期:2017-10-01 00:00:00
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journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
doi:10.1097/00042737-200505000-00016
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journal_title:European journal of gastroenterology & hepatology
pub_type: 临床试验,杂志文章
doi:10.1097/00042737-199802000-00008
更新日期:1998-02-01 00:00:00
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journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章,随机对照试验
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更新日期:2011-05-01 00:00:00
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journal_title:European journal of gastroenterology & hepatology
pub_type: 临床试验,杂志文章
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更新日期:1997-12-01 00:00:00
abstract:BACKGROUND:This study aimed to investigate the association between nonalcoholic fatty pancreas disease and the severity of acute pancreatitis (AP). PATIENTS AND METHODS:Among the 1662 AP patients admitted between August 2010 and August 2017, 82 eligible patients with moderately severe acute pancreatitis (SAP) and SAP ...
journal_title:European journal of gastroenterology & hepatology
pub_type: 杂志文章
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更新日期:2019-08-01 00:00:00
abstract::This leading article refers to the paper by Abdelrazeq AS, Owen C, Smith L, McAdam JG, Pearson HJ, Leveson SH. Nicorandil-associated para-stomal ulceration: case series Eur J Gastroenterol Hepatol 2006; 18:1293-1295. We apologise to all concerned for the dissociation between the two papers, which was due to an adminis...
journal_title:European journal of gastroenterology & hepatology
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更新日期:2007-05-01 00:00:00
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更新日期:2009-03-01 00:00:00