Development of a Mammalian suspension culture for expression of active recombinant human urokinase-type plasminogen activator.


:The development of specific catalytic inhibitors for the serine protease urokinase-type plasminogen activator (uPA) has been hindered due to difficulties in producing sufficient amounts of active recombinant uPA that is catalytically equivalent to native uPA. The purpose of this study was to develop an efficient system for the expression of recombinant human uPA that exhibits comparable proteolytic activity to that of the native protein. Since post-translational modifications (e.g. glycosylations) of uPA are necessary for efficient proteolytic activity, we have used a mammalian cell line [Chinese hamster ovary (CHO)-S] to express recombinant human uPA. CHO-S cells were selected to stably express full-length recombinant human uPA containing a hexahistidine tag at its C-terminus to permit purification by nickel-based affinity chromatography. Secretion of recombinant uPA into the culture media was confirmed by immunoblotting and the presence of an N-linked glycosylation was confirmed by PNGase sensitivity. Enzymatic activity of purified recombinant uPA was demonstrated using zymography and quantitatively compared to native uPA by kinetic analysis using an uPA-specific substrate. Native uPA and the recombinant uPA demonstrated comparable K(m) values (55.7 and 39 muM, respectively). Furthermore, inhibition studies using benzamidine resulted in a K(i) of 195 muM for native uPA, while recombinant uPA had a K(i) of 112 muM. These data indicate that recombinant human uPA expressed by CHO-S cells is functionally comparable to native uPA.






Atkins E,Zamora S,Candia BJ,Baca A,Orlando RA




Has Abstract


2005-09-01 00:00:00












  • Chromatid segregation analysis in native human lymphocyte anaphases using sequential fluorescence in situ hybridization.

    abstract::A sequential multiprobe fluorescence in situ hybridization technique was developed to study the 13, 18, 21, X and Y chromatid segregation in human lymphocytes anaphases cultures without antimitotic treatment. This method was used to know if exist any different chromosomes segregation in lymphocytes from Down syndrome ...


    pub_type: 杂志文章


    authors: Escudero T,Coll MD,Fuster C,Egozcue J

    更新日期:2002-09-01 00:00:00

  • Echinococcus granulosus protoscoleces promotes proliferation and invasion of hepatocellular carcinoma cells.

    abstract::There may exist a connection between Echinococcus granulosus infection and cancer development. Here, it is aimed to investigate specific effects of E. granulosus protoscoleces (PSCs) on the proliferation and invasion capacities of hepatocellular carcinoma (HCC) cells in vitro and ex vitro. HepG2 cells were cultured wi...


    pub_type: 杂志文章


    authors: Yasen A,Wang M,Ran B,Lv G,Aji T,Xiao H,Shao Y,Wen H

    更新日期:2021-02-01 00:00:00

  • The effect of glucose and glutamine on the intracellular nucleotide pool and oxygen uptake rate of a murine hybridoma.

    abstract::The effects of media concentrations of glucose andglutamine on the intracellular nucleotide pools andoxygen uptake rates of a murine antibody-secretinghybridoma cell line were investigated. Cells takenfrom mid-exponential phase of growth were incubated inmedium containing varying concentrations of glucose(0-25 mM) and...


    pub_type: 杂志文章


    authors: Barnabé N,Butler M

    更新日期:2000-10-01 00:00:00

  • Comparative cytogenetics between two species of the family Pseudopimelodidae (Siluriformes): occurrence of natural triploidy and supernumerary chromosomes.

    abstract::The Pseudopimelodidae family comprises 35 species however, cytogenetic studies have been performed in only six species. This study uncovered karyotypic data on Pseudopimelodus pulcher and Microglanis cottoides. Both species possessed 2n = 54, with 20m + 16sm + 10st + 8a and FN = 100 for P. pulcher and 30m + 14sm + 6st...


    pub_type: 杂志文章


    authors: Gouveia JG,de Moraes VP,Pires LB,da Rosa R,Dias AL

    更新日期:2015-03-01 00:00:00

  • Neuroscience of water molecules: a salute to professor Linus Carl Pauling.

    abstract::More than 35 years ago double Nobel laureate Linus Carl Pauling published a powerful model of the molecular mechanism of general anesthesia, generally referred to as the hydrate-microcrystal (aqueous-phase) theory. This hypothesis, based on the molecular behavior of water molecules, did not receive serious attention d...


    pub_type: 杂志文章


    authors: Nakada T

    更新日期:2009-04-01 00:00:00

  • Establishment of primary cell cultures from fish calcified tissues.

    abstract::Fishes have been recently recognized as a suitable model organism to study vertebrate physiological processes, in particular skeletal development and tissue mineralization. However, there is a lack of well characterized in vitro cell systems derived from fish calcified tissues. We describe here a protocol that was suc...


    pub_type: 杂志文章


    authors: Marques CL,Rafael MS,Cancela ML,Laizé V

    更新日期:2007-09-01 00:00:00

  • Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays.

    abstract::This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC50 and 2×EC50 values were 8 and 16 mg/ml concentrations...


    pub_type: 杂志文章


    authors: Ciğerci İH,Cenkci S,Kargıoğlu M,Konuk M

    更新日期:2016-08-01 00:00:00

  • Automated time-lapse microscopy and high-resolution tracking of cell migration.

    abstract::We describe a novel fully automated high-throughput time-lapse microscopy system and evaluate its performance for precisely tracking the motility of several glioma and osteoblastic cell lines. Use of this system revealed cell motility behavior not discernable with conventional techniques by collecting data (1) from cl...


    pub_type: 杂志文章


    authors: Fotos JS,Patel VP,Karin NJ,Temburni MK,Koh JT,Galileo DS

    更新日期:2006-05-01 00:00:00

  • Primary cell culture from embryos of the Japanese scallop Mizuchopecten yessoensis (Bivalvia).

    abstract::Primary cell cultures obtained from embryos of Mizuchopecten yessoensis (Bivalvia) survived for four months. Although the number of cells progressively decreased during the cultivation, mitotic cells were observed both at the first stages and at the end. A possibility of growing marine invertebrates cells in long term...


    pub_type: 杂志文章


    authors: Odintsova NA,Khomenko AV

    更新日期:1991-05-01 00:00:00

  • A rat mammary gland cancer cell with stem cell properties of self-renewal and multi-lineage differentiation.

    abstract::The cancer stem cell hypothesis posits that tumors are derived from a single cancer-initiating cell with stem cell properties. The task of identifying and characterizing cancer-initiating cells with stem cell properties at the single cell level has proven technically difficult because of the scarcity of the cancer ste...


    pub_type: 杂志文章


    authors: Cocola C,Sanzone S,Astigiano S,Pelucchi P,Piscitelli E,Vilardo L,Barbieri O,Bertoli G,Reinbold RA,Zucchi I

    更新日期:2008-09-01 00:00:00

  • Metabolic rates of vascular endothelial cellsin vitro.

    abstract::Cultures of endothelial cells and cell lines of endothelial origin were maintained at confluence without medium exchange for a period of 72 h. During this time period the concentration of nutrients - amino acids and glucose - and metabolic waste products - lactate and ammonium - was determined as well as cell vitality...


    pub_type: 杂志文章


    authors: Schrimpf G,Schröder M,Weitnauer E,Friedl P

    更新日期:1994-01-01 00:00:00

  • Features of free software packages in flow cytometry: a comparison between four non-commercial software sources.

    abstract::Flow cytometers designed to analyze large particles are enabling new applications in biology. Data analysis is a critical component of the process FCM. In this article we compare features of four free software packages including WinMDI, Cyflogic, Flowing software, and Cytobank. ...


    pub_type: 杂志文章


    authors: Sahraneshin Samani F,Moore JK,Khosravani P,Ebrahimi M

    更新日期:2014-08-01 00:00:00

  • Improvement in the antibody productivity of human-human hybridomas by transfection with Tac gene.

    abstract::The presence of a highly purified recombinant interleukin-2 (rIL-2) increased the production of immunoglobulin (IgM or IgG) by human-human hybridomas to 1.5-2.0 times the production by untreated cells. However, these cells did not react with anti-Tac (IL-2 receptor alpha) antibody. To enhance the response of the hybri...


    pub_type: 杂志文章


    authors: Iwamoto K,Shintani Y,Sasada R,Honjo T,Kitano K

    更新日期:1990-07-01 00:00:00

  • Sericin in the isolating solution improves the yield of islets isolated from the pancreas.

    abstract::Approximately half of the transplantable pancreatic islet tissue is lost during isolation, including the digestion and purification steps. Modifying the isolation method could increase the yield. This would enable the one donor-one recipient concept and improve the therapeutic effects of islet transplantation. This st...


    pub_type: 杂志文章


    authors: Yokoi S,Murakami M,Morikawa M,Goi T,Yamaguchi A,Terada S

    更新日期:2016-12-01 00:00:00

  • Engineering challenges in high density cell culture systems.

    abstract::High density cell culture systems offer the advantage of production of bio-pharmaceuticals in compact bioreactors with high volumetric production rates; however, these systems are difficult to design and operate. First of all, the cells have to be retained in the bioreactor by physical means during perfusion. The desi...


    pub_type: 杂志文章


    authors: Ozturk SS

    更新日期:1996-01-01 00:00:00

  • A direct computer control concept for mammalian cell fermentation processes.

    abstract::In the last 10 years, new assignments and the special demands of mammalian cells to the culture conditions caused the development of complex small scale fermentation setups. The use of continuous fermentation and cell retention devices requires appropriate process control systems. An arrangement for control and data-a...


    pub_type: 杂志文章


    authors: Büntemeyer H,Marzahl R,Lehmann J

    更新日期:1994-01-01 00:00:00

  • Liver stem cells.

    abstract::The capacity of hepatocytes and cholangiocytes to contribute to their own maintenance has long been recognized. More recently, studies have indicated the presence of both intra-hepatic and extra-hepatic stem/progenitor cell populations. The intraorgan compartment probably derives primarily from the biliary tree, most ...


    pub_type: 杂志文章


    authors: Theise ND

    更新日期:2003-03-01 00:00:00

  • Can macroscopic identification of large ovarian follicles be useful in eliminating follicles with impaired steroid function?

    abstract::"In vitro" studies, commonly using porcine ovarian follicles, may generate inconsistent results when atretic follicles are not eliminated from the pool of experimental follicles. The present experiment was conducted to test the practical value of the macroscopie identification of large porcine follicles, which were as...


    pub_type: 杂志文章


    authors: Dudek D

    更新日期:1994-01-01 00:00:00

  • Development and characterization of insect cell lines.

    abstract:CONCLUSIONS:With the wide availability of insect cell culture media, it can generally be considered a routine process to develop new cell lines. Exceptions to this statement do exist, of course. Difficulties may arise when attempting to culture a specific cell type. For example, while there are a few cell lines from in...


    pub_type: 杂志文章


    authors: Lynn DE

    更新日期:1996-01-01 00:00:00

  • Cytogenetic effects of nine Helichrysum taxa in human lymphocytes culture.

    abstract::Helichrysum Mill. (Asteraceae) species have been used in folk medicine for thousands of years in the world. The in vitro cytogenetic effects in human lymphocytes of nine Helichrysum taxa used in Turkey folk medicine were investigated. Blood samples were obtained from healthy donors, non-smoking volunteers, which were ...


    pub_type: 杂志文章


    authors: Eroğlu HE,Aksoy A,Hamzaoğlu E,Budak U,Albayrak S

    更新日期:2009-01-01 00:00:00

  • Analysis of toxicity effects of Di-(2-ethylhexyl) phthalate exposure on human bronchial epithelial 16HBE cells.

    abstract::Recent studies have indicated that Di-(2-ethylhexyl) phthalate (DEHP), the most commonly used plasticizer in daily-life products, could be dispersed in indoor air and induce human exposure via inhalation. DEHP has been reported to have effects on the respiratory system in both animal and human researches. The toxicity...


    pub_type: 杂志文章


    authors: Ma Y,Guo Y,Wu S,Lv Z,Zhang Q,Xie X,Ke Y

    更新日期:2018-02-01 00:00:00

  • Anti-photoaging activities of Sorbaria kirilowii ethanol extract in UVB-damaged cells.

    abstract::Sorbaria kirilowii (Regel) Maxim, a plant found in China, Korea, Japan, and east of Europe, is a common herb used for traditional medicinal purposes. However, its ability to prevent photoaging has not been studied. In this study, we investigated the anti-photoaging functions of an ethanol extract (Sk-EE) of S. kirilow...


    pub_type: 杂志文章


    authors: Chen H,Jang J,Kopalli SR,Yum J,Yoon K,Cho JY

    更新日期:2021-02-01 00:00:00

  • Efficient production of recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium.

    abstract::A fructose-based cell culture is suitable for the process control of protein production because of slow sugar consumption rate and low lactate accumulation. The fructose transporter, GLUT5, mediates its incorporation into cells and is required for the fructose-based culture. In order to produce efficiently recombinant...


    pub_type: 杂志文章


    authors: Inoue Y,Tsukamoto Y,Yamanaka M,Nakamura S,Inoue A,Nishino N,Kawahara H

    更新日期:2010-08-01 00:00:00

  • Distinct volume distribution of viable and non-viable hybridoma cells: A flow cytometric study.

    abstract::Light scattering properties of hybridoma cells were examined with flow cytometry. Viable and dead cells form two distinct populations. The distribution of the two populations changes during a batch culture. the concentration of dead cells measured by flow cytometry correlates well to that measured by hemacytometer. Th...


    pub_type: 杂志文章


    authors: Sen S,Srienc F,Hu WS

    更新日期:1989-05-01 00:00:00

  • Real-time transfer of lentiviral particles by producer cells using an engineered coculture system.

    abstract::Lentiviruses are quite effective gene delivery systems for stable production of genetically engineered human cells. However, prior to using lentivirus to deliver genetic materials to cells of interest, the normal course of production of these lentiviruses involves a lengthy collection, purification, preservation, and ...


    pub_type: 杂志文章


    authors: Timmins LM,Patel RS,Teryek MS,Parekkadan B

    更新日期:2019-10-01 00:00:00

  • Mechanisms of resistance to alkylating agents.

    abstract::Alkylating agents are the most widely used anticancer drugs whose main target is the DNA, although how exactly the DNA lesions cause cell death is still not clear. The emergence of resistance to this class of drugs as well as to other antitumor agents is one of the major causes of failure of cancer treatment. This pap...


    pub_type: 杂志文章


    authors: Damia G,D'Incalci M

    更新日期:1998-09-01 00:00:00

  • Silencing of soluble epoxide hydrolase 2 gene reduces H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells via activating PI3K/Akt/GSK3β signaling pathway.

    abstract::Oxidative stress plays a vital role in the occurrence and development of intestinal injury. Soluble epoxide hydrolase 2 gene (EPHX2) is a class of hydrolytic enzymes. We aim to explore the effects and molecular mechanism of siEPHX2 on H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells. IEC-6 cells ...


    pub_type: 杂志文章


    authors: Li J,Luo J,Zhang Y,Tang C,Wang J,Chen C

    更新日期:2020-02-01 00:00:00

  • Anin vitro model for the biological evaluation of dental materials using human periodontal ligament cells.

    abstract::Anin vitro system is described which uses human diploid cells derived from the periodontal ligament. It simulates the clinical situation and is suitable for rapid cytotoxicity screening of dental materials. It can also be adapted to study various factors influencing biocompatibility. ...


    pub_type: 杂志文章


    authors: Adams AM,Soames JV,Searle RF

    更新日期:1988-02-01 00:00:00

  • Effects of cell density and phorbol esters on the expression of epidermal growth factor receptors.

    abstract::Previously, it has been shown that the binding of epidermal growth factor (EGF) by a wide range of cells decreases as cell density increases. In this report, we demonstrate that KB cells treated chronically with phorbol esters continue to exhibit decreases in EGF receptor binding as cell density increases. This findin...


    pub_type: 杂志文章


    authors: Rizzino A,Huebert C,Kuszynski C,Wilder PJ

    更新日期:1991-10-01 00:00:00

  • Apoptosis-inducing activities of Halopteris scoparia L. Sauvageau (Brown algae) on cancer cells and its biosafety and antioxidant properties.

    abstract::The aim of this study was to reveal the biological activities and in vivo toxicity profiles of n-hexane, chloroform and methanol extracts of brown algae Halopteris scoparia L. Sauvageau. In this study, extracts were tested for their phytochemical contents and antioxidant activities. The cytotoxic activities of the ext...


    pub_type: 杂志文章


    authors: Güner A,Nalbantsoy A,Sukatar A,Karabay Yavaşoğlu NÜ

    更新日期:2019-06-01 00:00:00