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Cultivation of human liver cell lines with microcarriers acting as biological materials of bioartificial liver 总被引:3,自引:0,他引:3
Subjectheadingsbioartificialliver;livercelllines;microcarrierINTRODUCTIONTheanimalexperimentsofextracorporealbioartificialliversuggestedthatthedevicecouldprovidespecialassistancetohepaticfunctions,andtheeffectsofitsprimaryclinicalapplicationwas.1~3]encouraging.Althoughfewsuccessfulstudieswerereportedonhumancelllineactingasthebiologicalmaterialofbioartificialliver,itisratherconspicuous["'j9andhasopenedupanewpathforthestudyofbioartificialliver.Tomeettheprincipalneedsofbioartificialliverfunc… 相似文献
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Ishii Y Saito R Marushima H Ito R Sakamoto T Yanaga K 《World journal of gastroenterology : WJG》2008,14(17):2740-2747
AIM: To examine the efficacy of the radial flow bioreactor (RFB) as an extracorporeal bioartificial liver (BAL) and the reconstruction of liver organoids using embryonic pig liver cells. METHODS: We reconstructed the liver organoids using embryonic porcine liver cells in the RFB. We also determined the gestational time window for the optimum growth of embryonic porcine liver cells. Five weeks of gestation was designated as embryonic day (E) 35 and 8 wk of gestation was designated as E56. These cells were cultured for one week before morphological and functional examinations. Moreover, the efficacy of pulsed adminisbation of a high concentration hepatocyte growth factor (HGF) was examined. RESULTS: Both cell growth and function were excellent after harvesting on E35. The pulsed administration of a high concentration of HGF promoted the differentiation and maturation of these fetal hepatic cells. Microscopic examination of organoids in the RFB revealed palisading and showed that bile duct-like structures were well developed, indicating that the organoids were mini livers. Transmission electron microscopy revealed microvilli on the luminal surfaces of bile duct-like structures and junctional complexes, which form the basis of the cytoskeleton of epithelial tissues. Furthermore, strong expression of connexin (Cx) 32, which is the main protein of hepatocyte gap junctions, was observed. With respect to liver function, ammonia detoxification and urea synthesis were shown to be performed effectively.
CONCLUSION: Our system can potentially be applied in the fields of BAL and transplantation medicine. 相似文献
CONCLUSION: Our system can potentially be applied in the fields of BAL and transplantation medicine. 相似文献
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目的通过对肝细胞癌(HCC)中3种耐药基因MDR1、MRP、GST-π的检测,探讨肝细胞癌中耐药基因的表达特点及临床意义。方法应用逆转录-聚合酶链反应(RT-PCR)技术检测3种耐药基因在51例肝细胞癌组织和10例正常肝组织中的表达。结果 (1)MDR1、MRP、GST-π在肝细胞癌中的表达分别为0.55±0.27、0.62±0.29、0.64±0.31,正常肝组织中的表达分别为0.23±0.10、0.25±0.07、0.26±0.12,耐药基因在肝细胞癌中的表达高于正常肝组织,差异具有统计学意义(P〈0.05);(2)耐药基因的表达与肿瘤Edmondson分级呈正相关(P〈0.05);(3)MRP与GST-π的表达相关。结论肝细胞癌中存在有原发性耐药的现象,且多种机制并存。MDR1、MRP、GST-π在肝细胞癌中有较高的表达。联合检测对制定科学有效的治疗方案有一定价值。 相似文献
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Establishment and characterization of four human hepatocellular carcinoma cell lines containing hepatitis B virus DNA 总被引:18,自引:1,他引:17
Lee JH Ku JL Park YJ Lee KU Kim WH Park JG 《World journal of gastroenterology : WJG》1999,5(4):289-295
INTRODUCTIONHepatocelularcarcinoma(HCC)isoneofthemostprevalentmalignantdiseasesencounteredintheworld,kilingupto1milionpeoplea... 相似文献
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Developments in bioartificial liver research: concepts, performance, and applications 总被引:5,自引:0,他引:5
As an alternative to liver transplantation, numerous researchers have been working toward the goal of development of a fully
functional artificial liver. In recent years, artificial liver support systems have been advocated as interim treatments for
patients awaiting hepatocyte replacement therapy or liver transplantation; so-called "bridging" treatments. It is recognized
that an effective artificial liver system requires: (1) a viable and highly functional hepatocyte cell line, (2) a suitable
bioreactor environment and peripheral control systems, and (3) an effective extracorporeal circulatory system to incorporate
an artificial liver system. Conventional systems have, however, suffered from various drawbacks, including incompatibility
of cell cultures derived from non-human cells, insufficient cell proliferation, rapid deterioration of cellular function due
to an impoverished cellular environment, and lack of system scalability. A newly established artificial liver system overcomes
many of these problems and demonstrates a long-term capacity to maintain multiple liver-specific functions, such as protein
synthesis, enzyme activity, and drug metabolism, both quantitatively and qualitatively. The present review provides an overview
of the concepts underpinning artificial liver systems, the performance of presently available systems and the practical applications
of available systems and those in development.
Received: February 7, 2000 / Accepted: February 25, 2000 相似文献
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目的研究mdm2基因在原发性肝细胞癌(HCC)中的表达并探讨其与p53基因突变的关系.方法用银染PCRSSCP法检测p53基因第5~8外显子的突变,原位杂交检测mdm2基因mRNA的表达,SABC法检测mdm2蛋白的表达.结果393%(11/28)的病例有异常的电泳迁移率.p53基因突变与肿瘤的大小、分化及转移无关.原位杂交显示9例HCC出现mdm2基因mRNA增加,7例HCC可检测到mdm2蛋白表达,mdm2基因表达与HCC的大小、分化及是否转移无关.Ⅰ~Ⅱ级HCC中mdm2阳性表达率(133%)明显低于Ⅲ~Ⅳ级HCC中的阳性表达率(538%).11例有p53基因突变的HCC中,只有3例出现mdm2基因表达,另外6例有mdm2过表达的HCC未见p53基因突变.p53基因突变的HCC与p53基因无突变的HCC相比,mdm2基因表达阳性率无显著差别.结论p53基因突变和mdm2基因表达在原发性HCC的发病中起重要作用.mdm2基因表达与HCC的恶性程度相关.mdm2基因表达与p53基因是否突变无关. 相似文献
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抑癌基因PTEN及p53在肝细胞肝癌中表达的免疫组化研究 总被引:2,自引:0,他引:2
为探讨肝细胞肝癌组织中抑癌基因PTEN及p53蛋白的表达情况及临床病理意义。应用免疫组织化学技术检测了41例肝细胞肝癌及其相应的癌旁组织中PTEN和p53蛋白的表达情况。41例癌旁组织PTEN全部阳性表达,肝细胞肝癌组织中PTEN阳性表达率39%,阳性信号显示于胞浆中。p53阳性表达率51%,PTEN蛋白在肝细胞肝癌组织中的阳性表达与组织分化程度明显相关,高分化癌的阳性率为73%,低分化癌阳性率27%。肝细胞肝癌细胞中存在较高比例的PTEN蛋白阴性表达,说明在肝细胞肝癌的发生发展中PTEN基因失活起着重要作用,它的阳性表达可能有一定的预后意义。 相似文献
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目的观察腺病毒介导的Runx3基因(Ad-Runx3)联合顺铂(DDP))对肝癌细胞HLE增殖的抑制效果。方法通过Ad-Runx3感染HLE,采用RT-PCR方法检测Runx3基因在HLE中的转录,用Ad-Runx3联合DDP处理培养的HLE,采用Western blot法检测HLE中Runx3表达情况,MTT法检测HLE细胞生长抑制率,流式细胞术检测HLE细胞周期和凋亡率。结果 Ad-Runx3感染HLE后即有目的基因转录,感染48 h后,Runx3呈高表达;加入100感染复数(MOI)的Ad-Runx3与6.25μg/ml的DDP后5 d,HLE细胞生长抑制率达92.46%±1.13%,显著高于单加Ad-Runx3者的59.28%±1.37%和单加DDP者的46.37%±2.51%(P均〈0.05)。加入Ad-Runx3与DDP的HLE其细胞阻滞于G2/M期,S期细胞减少,细胞凋亡率为18.62%±2.48%,显著高于单用Ad-Runx3者的8.66%±0.78%和DDP者的7.48%±0.32%(P均〈0.05)。结论 Ad-Runx3可增强DDP致肝癌细胞HLE增殖抑制作用,提高其细胞凋亡率。 相似文献
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5-脱氧杂氮胞苷抑制肝癌细胞株生物学行为的机制 总被引:4,自引:3,他引:1
目的 DNA甲基化模式的改变伴随着永生性细胞系的确立,生长凋控基因CpG岛的重新甲基化可能导致了其转录的不活跃,在体外培养的情况下,使得肿瘤细胞选择了有利于生长的条件.我们用DNA甲基转移酶抑制剂5-脱氧杂氮胞苷处理两株肝癌细胞,研究其对肝癌细胞的影响,探讨DNA甲基化异常与肝细胞癌间的相关性及5-脱氧杂氮胞苷对肝癌细胞株恶性生物学行为的影响及其机制。方法 用5-脱氧杂氮胞苷处理肝癌细胞株SMMC-7721和HePG2,然后采用相差显微镜观察药物处理前后细胞的形态变化,采用MTT法观察细胞的生长速度变化,采用流式细胞仪检测细胞周期、细胞凋亡率、P16蛋白表达的变化.采用RT-PCR法比较p16和甲基转移酶mRNA表达量的变化,比较裸鼠致瘤性的大小。结果 5-脱氧杂氨胞苷处理后细胞形态趋于规则,生长速度减慢.SMMC-7721细胞,G1期细胞增加了8.5%,而s期和G2/M期细胞分别减少了47.8%和10.4%.HePG2细胞,G1期细胞增加了3.5%,S期减少了46.2%.G2/M期增加了23.7%.两细胞凋亡率分别增加了91.6%和133.3%.P16蛋白表达分别增加了23.4%和20.9%,p16mRNA表达增高,而DNA甲基转移酶mRNA表达明显降低,裸鼠皮下移植瘤生长减慢。结论 肝细胞癌的发生与DNA甲基化异常有关,甲基化酶抑制剂5-脱氧杂氮胞苷可能通过抑制甲基转移酶抑制抑癌基因启动子区域的高甲基化,恢复其生长调控功能,从而使肝癌细胞株的恶性生物学行为发生逆转。 相似文献
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CAO Guang Wen GAO Jun DU Ping QI Zhong Tian KONG Xian Tao 《World journal of gastroenterology : WJG》1997,3(3):139-142
ConstructionofretroviralvectorstoinduceastrongexpresionofhumanclasⅠinterferongeneinhumanhepatocelularcarcinomacelsinvitroCAOG... 相似文献
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Recurrence of hepatocellular carcinoma as a mixed hepatoblastoma after liver transplantation 总被引:1,自引:0,他引:1
BACKGROUND: Hepatoblastoma is an exceptional cause of primary malignant liver tumour in the adult. PATIENT: The case is reported of an adult patient transplanted for alcoholic cirrhosis complicated by multifocal hepatocellular carcinoma in whom a recurrence in the form of a mixed hepatoblastoma invading the whole transplanted liver developed three months after liver transplantation. METHODS: Complete clinical, histopathological, and immunohistochemical data were reviewed. RESULTS: The recurrent tumour invaded the whole liver. The major component was a mixed hepatoblastoma, with an epithelial component expressing cytokeratin and a mesenchymal component expressing vimentin. The tumour also contained a minor hepatocarcinomatous component expressing alpha fetoprotein. The rapid growth of the tumour prevented any attempt at treatment. Although direct evidence is lacking, the most likely hypothesis to explain the observations is a marked phenotypic change in the initial malignant population at recurrence. CONCLUSION: This case supports a possible filiation between hepatocellular carcinoma and hepatoblastoma in adults. 相似文献
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SEIICHI USUDA KANAME YOSHIZAWA KOJI YABU KENDO KIYOSAWA 《Journal of gastroenterology and hepatology》1993,8(6):517-523
We performed a detailed analysis of immune responses in a hepatocellular carcinoma (HCC) cell line and effector cells obtained from a patient with HCC. We examined the cytotoxic activity of natural killer (NK) cells, lymphokine-activated killer (LAK) cells and cytotoxic T lymphocytes (CTL) against an autologous tumour cell line (SUHC-1) to investigate the immune mechanism of human lymphocytes against HCC cells. Cytotoxic T lymphocytes were induced by co-culturing of peripheral blood lymphocytes (PBL) and SUHC-1 cells, mixed lymphocyte and tumour cell culture (MLTC). The susceptibility of SUHC-1 to NK and LAK cells was similar to that of other allogeneic cell lines, such as K562, PLC/PRF/5 and Mahlavu. Effector cells induced in the primary MLTC had high cytotoxic acitivity but were not specific for SUHC-1. Cytotoxic T lymphocytes with specific activity against SUHC-1 were induced after PBL were stimulated five times at 7–10 day intervals with SUHC-1 and low-dose recombinant interleukin-2 (rIL-2), suggesting that as the culture progressed, broadly reactive effector cells disappeared and specific effector cells survived. The specific effector cells were identified as CD3+/CD4+ and CD+/CD8+ T-lymphocyte subsets. The recognition mechanisms of CD3+/CD4+ CTL remain unresolved because the cytotoxicities were not inhibited by anti-CD4 and anti-major histocompatibility complex (MHC) class II monoclonal antibodies (MoAb). Treatment of cells with anti-CD3, anti-CD8 and anti-MHC class I MoAb partially inhibited lysis. These results demonstrated that the T-cell receptor (TCR)/CD3 complex appeared to be involved in SUHC-1 specific antigen recognition and antigen recognition of CD3+/CD8+ CTL was MHC class I restricted. 相似文献
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目的:应用基因芯片技术研究原发性肝细胞癌组织中的差异基因表达谱改变,以寻找肝细胞癌相关基因。方法:抽提正常肝组织和肝癌组织中的mRNA来制备探针,经杂交、洗涤后,通过计算机扫描分析正常肝组织和肝癌组织基因表达谱的差异情况。结果:在10000个候选基因中,筛选出102条差异表达基因,表达上调的有42条,表达下调的有60条。未知基因12条。结论:基于DNA微阵矩技术的肿瘤基因表达谱分析能够高通量筛选与肝癌发生发展相关的基因。 相似文献
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不同转移潜能人肝癌HCCLM3、MHCC97-L细胞系血管生成拟态基因的表达 总被引:1,自引:0,他引:1
目的采用细胞外基质与粘连分子Real-time PCR基因芯片筛选高、低转移潜能人肝癌HCCLM3、MHCC97-L细胞系差异表达的血管生成拟态基因,探讨血管生成拟态的机制。方法建立HCCLM3、MHCC97-L三维培养体系,倒置显微镜下观察其血管样结构形成情况。细胞培养24 h后,采用细胞外基质与粘连分子Real-timePCR基因芯片对HCCLM3、MHCC97-L的表达基因进行筛选。结果培养24 h,HCCLM3形成的血管样结构较MHCC97-L的长(P〈0.01)。筛选出20个差异基因,其中CD44、COL6A1、COL4A2、ECM1、ITGA1、ITGA3、MMP1、MMP2、SPP1、TNC、COL6A2、MMP7和MMP10共13个基因在HCCLM3中的表达较MHCC97-L明显上调,CTNND2、COL12A1、COL14A1、ITGAL、TIMP3、MMP16和VTN共7个基因明显下调。结论高、低转移潜能人肝癌HCCLM3、MHCC97-L细胞系体外形成血管生成拟态有差异,其原因可能与HCCLM3差异表达某些细胞外基质和粘连分子相关基因有关。 相似文献
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Yutaka Yata Feng Xue Terumi Takahara Hiroshi Kudo Katsuharu Hirano Satoshi Yasumura Masami Minemura Andrew E Scanga Toshiro Sugiyama 《Hepatology research》2010,40(3):304-310
Background: Current chemotherapy for advanced hepatocellular carcinoma (HCC) is insufficient; only sorafenib has been proven to provide a modest survival benefit. A future direction of chemotherapy is to tailor treatment based on the chemosensitivity of each individual tumor. By doing so, only patients who stand to benefit from therapy will be exposed to potential side‐effects and morbidity. Although the use of docetaxel (DTX) for the treatment of lung, breast and gastric cancer has been reported, there are few reports about its use in the setting of HCC. Methods: To examine the efficacy of DTX for HCC, we established a human hepatoma cell line (TK cell) from the patient's malignant ascites from peritoneal carcinomatosis and treated it with DTX in vitro. Results: After we confirmed the efficacy of DTX in vitro, we treated our patient with DTX with positive results. Conclusion: In this study, we present a therapeutic approach by using DTX that supports the potential usefulness of personalized medicine in vitro and demonstrates it clinically. 相似文献