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Establishment and characterization of a rat pancreatic stellate cell line by spontaneous immortalization

World Journal of Gastroenterology 2003年第12期9卷 2751-2758页

作者： Atsushi Masamune Masahiro Satoh Kazuhiro Kikuta Noriaki Suzuki Tooru Shimosegawa Division of Gastroenterology Tohoku University Graduate School of MedicineSendaiJapan A.M.and M.S.equally contributed to this work

AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous ***: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift *** of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme ***: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely *** cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP ***-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP ***: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.

关键词： Animals Base Sequence cell Culture Techniques cell line, transformed cells, Cultured Cystic Fibrosis Cytoskeletal Proteins Extracellular Matrix Proteins Immunohistochemistry NF-kappa B Oligonucleotide Probes Pancreas Rats Research Support, Non-U.S. Gov't Transcription Factor AP-1

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Transplantation of human hepatocytes into tolerized genetically immunocompetent rats

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World Journal of Gastroenterology 2001年第3期7卷 324-330页

作者： EdwinC.Ouyang CatherineH.Wu CherieWalton KittichaiPromrat GeorgeY.Wu Department of Medicine Division of Gastroenterology-HepatologyUniversity of Connecticut Health CenterRm. AM-045263 Farmington AvenueFarmingtonCT 06030 Correspondence to:George Y.Wu.M.D. Ph.D.Department of Medicine.Division of Gastroenterology-Hepatology.University of Connecticut I lealth CenterRm.AM-045.263 Farmington Avenue.FarmingtonC T 06030

AIM: To determine whether normal genetically immunocompetent rodent hosts could be manipulated to accept human hepatocyte transplants with long term survival without immunosuppression. METHODS: Tolerance towards human hepatocytes was established by injection of primary human hepatocytes or Huh7 human hepatoma cells into the peritoneal cavities of fetal rats. Corresponding cells were subsequently transplanted into newborn rats via intrasplenic injection within 24h after birth. RESULTS: Mixed lymphocyte assays showed that spleen cells from non-tolerized rats were stimulated to proliferate when exposed to human hepatocytes, while cells from tolerized rats were not. Injections made between 15 d and 17 d of gestation produced optimal tolerization. Transplanted human hepatocytes in rat livers were visualized by immunohistochemical staining of human albumin. By dot blotting of genomic DNA in livers of tolerized rats 16 weeks after hepatocyte transplantation, it was found that approximately 2.5 X 10(5) human hepatocytes survived per rat liver. Human albumin mRNA was detected in rat livers by RT-PCR for 15 wk, and human albumin protein was also detectable in rat serum. CONCLUSION: Tolerization of an immuno-competent rat can permit transplantation, and survival of functional human hepatocytes.

关键词： Albumins Animals cell line, transformed Disease Models, Animal Female Gene Expression Graft Survival Hepatitis Hepatoblastoma Hepatocytes Humans Immune Tolerance Immunocompetence Liver Liver Neoplasms Lymphocyte Culture Test, Mixed Microscopy, Confocal Pregnancy RNA, Messenger Rats Rats, Sprague-Dawley Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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