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Static three-dimensional culture of human hepatocarci-noma cell with microcarriers

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Chinese Science Bulletin 2001年第20期46卷 1704-1709页

作者： Zhang Jianmin Wang Hongfang feng meifu Chinese Acad Sci State Key Lab Biomembrane & Membrane Biotechnol Inst Zool Beijing 100080 Peoples R China

Traditional monolayer culture is still the common method used in hepatocarcinoma cell culture in vitro. For lack of the structure similar to that in vivo, it is disadvantageous in the research of the relation between structure and function. We established a three-dimensional (3-D) culture model of human hepatocarcinoma cell (BEL-7402) in vitro by using microcarrier cytodex-3 in static condition. The results of SEM, TEM, enzyme activity and flow cytometry indicated that the cells were polygonal-shaped and arranged in multilayers. Intercellular space was 0.5-2.0 (Am wide where lots of microvilli could be seen. Adjacent cells were connected with desmosomes and localized membrane projects. More than 90% of the cells were viable and maintained the consumption of glucose and the expression of EGF receptor. The intracellular ALT, AST and LDH-L activities were higher in 3-D culture than those of monolayer culture. Compared with monolayer culture, this 3-D culture with the structure similar to trabecular

关键词： microcarrier three-dimensional culture hepatocarci- noma BEL-7402 model.

Genistein inhibits human TNF-α-induced porcine endothelial cell adhesiveness for human monocytes and natural killer cells

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Chinese Science Bulletin 2002年第6期47卷 470-475页

作者： Xiaofeng Zhang Yan Gu Zhimin feng Liqin Ban meifu feng National Laboratory of Biomembrane and Membrane Biotechnology Institute of Zoology Chinese Academy of Sciences Beijing China

Cellular immune response is a major barrier to xenotransplantation. Human tumor necrosis factor-α (hTNF-α) possesses cross-species activity and directly amplifies the immune rejection via the upregulation of adhesion molecules on porcine endothelium. We investigated the role of protein tyrosine phosphorylation in the induction of expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1), and the augmentation of adhesion of human peripheral blood monocytes (PBMo) and natural killer cells (PBNK), after rhTNF-α-stimulation of porcine aortic endo-thelial cells (PAEC) in vitro. rhTNF-α-increased adhesiveness of PAEC for both PBMo and PBNK was dose-dependently reduced by pretreatment of PAEC with the selective protein tyrosine kinase (PTK) inhibitor genistein. The inhibitory effect occurred at the early time of PAEC activation triggered by rhTNF-α, and was completely reversible. PTK activity assay indicated that genistein also suppressed rhTNF-α stimulated activation of protein tyrosine

关键词： genistein human tumor necrosis factor-α (hTNF-α) porcine aortic endothclial cells (PAEC) human peripheral blood monocytes (PBMo) human peripheral blood natural killer cells (PBNK) cell adhesion.

Adhesion of subsets of human blood mononuclear cells to porcine endothelial cells

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Chinese Science Bulletin 2000年第7期45卷 626-630页

作者： Xiaofeng Zhang Zhimin feng meifu feng Hongfang Wang Ban Liqin Chinese Acad Sci Inst Zool State Key Lab Biomembrane & Membrane Biotechnol Beijing 100080 Peoples R China

Cellular immune response is a major barrier to xenotransplantation, and cell adhesion is the first step in intercellular recognition. Flow-cytometric adhesion assay has been used to investigate the differential adhesions of monocyte (Mo), natural killer celi (NK) and T lymphocyte (T) present within human peripheral blood mononuclear cells (PBMC) to porcine aortic endothelial cells (PAEC), and to demonstrate the effect of human interferon-γ (hIFN-γ) or/and tumor necrosis factor-α (hTNF-α) pretreatment of PAEC on their adhesiveness f or different PBMC subsets. The preferential sequence for PBMC subset binding to resting PAEC is Mo, NK and T cells, among which T cells show the slightest adherence; hTNF- a can act across the species, and augment Mo, NK and T cell adhesion ratios by 40%, 110% and 3 times, respectively. These results confirm at the cell level that host Mo and NK cells are major participants in the cellular xenograft rejection, thereby, providing a prerequisite for further studying the

关键词： human peripheral blood mononuclear cell (PBMC) porcine aortic endothelial cell (PAEC) human tumor necrosis factor-α (hTNF-α) cell adhesion phenotypic analysis.

Phenotypic analysis of the medullary-type CD4^-CD8^+ thymocytes in mice

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Chinese Science Bulletin 2000年第3期45卷 232-237页

作者： TIAN Tian WANG Hongfang feng meifu CHEN Weifeng Department of Immunology Beijing Medical UniversityBeijing 100083China Institute of Zoology Chinese Academy of SciencesBeijing 100080China

Phenotypic analysis of the medullary-type CD4 CD8+ (CD8SP) thymocytes has revealed phenotypic heterogeneity within this cell population. The phenotype of mature peripheral CD8+T cells is TCRαβ+CD3+Qa-2+HSA3G11 6C10 , whereas in the medullary-type CD8SP thymocytes, 20% are Qa-2+; 33%, HAS ; 30%, 3G11 ; and 70% are 6C10 . The disparate expression patterns of these four cell surface markers suggest that medullary-type CD8SP thymocytes may undergo phenotypic maturation process. According to the distribution of these four celi surface markers, six subgroups of CD8SP thymocytes have been identified. The precursor-progeny relationship along with developmental pathway is postulated as follows: 6C10+HSA+3G11 Qa-2 - 6C10+HSA+ 3G11+Qa-2 6C10 HSA+3G11+Qa-2 -6C10 HSA 3G11 +Qa-2 -6C10 HSA 3G11 Qa-2 -6C10 HA S 3G11 Qa-2+, the cells inthe last subgroup exit the thymus and home into periphery.

关键词： CD^4 CD8^+ medullary-type thymocytes cell phenotype phenotypic differentiation.

Progress on Microgravity Sciences in China

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空间科学学报 2006年第Z1期26卷 150-159页

作者： LIU Qiusheng NIE Yuxin feng meifu HU Wenrui Institute of Mechanics National Microgravity Laboratory Chinese Academy of Sciences Beijing 100080 China Institute of Physics Chinese Academy of Sciences Institute of Zoology Chinese Academy of Sciences Institute of Mechanics National Microgravity Laboratory Chinese Academy of Sciences Beijing 100080 China

The main progress of the research activities on microgravity fluid physics, combustion, biotechnology research and fundamental Physics in China are briefly summarized in the present paper. The major space missions and experimental results obtained on board the Chinese recoverable/nonrecoverable satellites and the Chinese manned spaceship named "Shen Zhou" are presented summarily. The recent main activities of the ground-based studies in China are introduced in brief.

关键词： Microgravity sciences, Fluid physics, Combustion, Fundamental physics, Biotechnology

Human endothelial senescence can be induced by TNF-α

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Chinese Science Bulletin 2002年第2期47卷 118-122页

作者： Jianjun Zhou Yunfei Gao Hongfang Wang Quan Chen Caihong Wu meifu feng The Natl. Key Lab. of Biomembrane College of Life Sciences Peking University Beijing 100871 China

TNF-α is one of the most important proin-flammatory cytokines in mediating multiple physio-patho-logical functions during immunological responses. Vascular endothelial cells, when stimulated by TNF-α, can increase the expression of multiple cytokines and cellular adhesion molecules and, in turn, actively promote the inflammatory responses by recruiting and activating of leukocytes to the inflammatory site. In addition to endothelial death induced by TNF-α, we found for the first time that TNF-α can also induce the human endothelial cells senescence. The induced senescent endothelial cells will display SA-B-Gal staining and they were arrested in G0-G1 phase. We found that △(?)m would always be up-regulated in response to TNF-α stimula-tion at early time but when the cells become senescent, △(?)m shows a tendency to decrease. It may reflect the sthenic func-tion of mitochondria at early time in response to TNF-α stimulation and decay when the endothelial cells were in-duced senescent. ROS

关键词： TNF-α endothelial cells inflammation senescence.

Advances in studies on hepatic stem cells

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Progress in Natural Science:Materials International 2003年第3期 8-14页

作者： HE Zuping, TANG Yuefeng, LIU Yabing and feng meifu(State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China) State Key Laboratory of Biomembrane and Membrane Biotechnology Institute of Zoology Chinese Academy of Sciences Beijing 100080 China

The question whether hepatic stem cells exist or not has been debated for several decades. Current researches confirmthat there are hepatic stem cells in the liver. Oval cells, putative bipotential hepatic stem cells, are probably located within canals of Her-ing, portal tracts or branches of biliary trees. Bone marrow is a potential source of oval cells, indicating that there exists a close relationshipbetween liver and hematopoiesis in adulthood. Hepatic stem cells are able to proliferate in vitro and can be induced to differentiate intohepatocytes. This will provide a promising approach of cell transplantation, tissue engineering and gene therapy for liver diseases. In thisreview, the evidence of their presence, origin, identification, proliferation in vitro, differentiation by induction, application prospects ofhepatic stem cells and future directions for the field are discussed.

关键词： hepatic stem celis origin identification proliferation in vitro differentiation by induction clinical application.

GM-CSF-induced Ca_(2+) -activated non-voltage-dependent inwardly rectifying K_+ channel in murine peritoneal exudate macrophages

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Chinese Science Bulletin 1997年第11期42卷 947-952页

作者： ZHANG Xiaofeng, feng meifu, WU Caihong and ZHOU Pei’aiState Key Laboratory of Biomembrane and Membrane Biotechnology, Chinese Academy of Sciences, Beijing 100080, China 1 . In-stitute of Zoology, Chinese Academy of Sciences, Beijing 100080, China 2 College of Life Sciences, Peking University, Beijing 100871, China State Key Laboratory of Biomembrane and Membrane Biotechnology Chinese Academy of Sciences Beijing China Institute of Zoology Chinese Academy of Sciences Beijing China College of Life Sciences Peking University Beijing China

GRANULOCYTE-MACROPHAGE colony-stimulating factor(GM-CSF),a member in the family ofglycoprotein cytokines,can not only act to stimulate proliferation of immature progenitors,butalso enhance differentiated functions of mature effector *** has two kinds of biologicaleffects on mature effector cells,*** effect and indirect *** latter is also

关键词： K+ channel patch clamp GM-CSF macrophage.

Enrichments identification and differentiation of hepatic stem/progenitor cells derived from human fetal liver

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Enrichments identification and differentiation of hepatic st...