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991.
Hemophilia A is one of the major inherited bleeding disorders caused by a deficiency or abnormality in coagulation factor VIII (FVIII). Hemophiliacs have been treated with whole plasma or purified FVIII concentrates. The risk of transmitting blood-borne viruses and the cost of highly purified FVIII are the major factors that restrict prophylaxis in hemophilia therapy. One of the challenges created by the biotechnology revolution is the development of methods for the economical production of highly purified proteins in large scales. Recent developments indicate that manipulating milk composition using transgenesis has focused mainly on the mammary gland as a bioreactor to produce pharmaceuticals. In the present study, a hybrid gene containing bovine -lactalbumin and human FVIII cDNA was constructed for microinjection into the pronuclei of newly fertilized mouse eggs. The LA-hFVIII hybrid gene was confirmed to be successfully integrated and stably germ-line transmitted in 12 (seven females/five males) lines. Western-blot analysis of milk samples obtained from eight of the transgenic founders and F1 offspring indicated that the recombinant hFVIII was secreted into the milk of the transgenic mice. The concentrations of rFVIII ranged from 7.0 to 50.2g/ml, over 35–200-fold higher than that in normal human plasma. Up to 13.4U/ml of rFVIII was detected in an assay in which rFVIII restored normal clotting activity to FVIII-deficient human plasma. 相似文献
992.
Ralf Baumeister 《Briefings in Functional Genomics and Prot》2002,1(1):53-65
The completion of the genome sequences of several model organisms and the recent development of high throughput procedures to map genes, expression patterns and interactions is providing a steadily increasing number of candidate target genes. The function of most of these genes still remains unknown. Therefore, there is a growing demand in genetically tractable animal models in which the function of individual factors can be studied in large scale, particularly of those that are thought to segregate with human disorders. In this paper, current methods to validate target gene function and the advantages of different model organisms are compared. 相似文献
993.
本文概述了实验动物的分类、特点及其在医学生物学中的应用。在医学生物学的发展过程中,实验动物的重要性已愈来愈被人们所认识。合理地选用实验动物对达到预定的实验目的起着至关重要的作用。 相似文献
994.
《Experimental parasitology》1998,90(3):270-276
Breijo, M., Spinelli, P., Sim, R.B., and Ferreira, A. M. 1998.Echinococcus granulosus:An intraperitoneal diffusion chamber model of secondary infection in mice.Experimental Parasitology90, 270–276. The present work describes a new experimental model of secondary infection which allows, through the recovery of the parasite together with its localin vivoenvironment, examination of the local nonadaptive immune response of the infected host and the differentiation of the parasite from protoscoleces to cysts. In this model we administered protoscoleces within silicone diffusion chambers, previously implanted into the peritoneal cavities of mice. The process of designing the model involved, first, determination of the optimal time postimplantation to infect the mice and, second, evaluation of the parasite's ability to establish infection within the chambers. The optimal time for infection was considered to be after the inflammation caused by implantation of the chambers had subsided. Our results showed that by day 20 postsurgery, three parameters used as indications of inflammation (complement C3, serum amyloid P protein, and polymorphonuclear cells in the peritoneum and in the chamber contents) had reverted to their normal levels. In our study of parasite differentiation, we found that 2–3% of the total number of parasites inoculated into the chambers were recovered as viable cysts after 100 days. Throughout the infection period, the population of parasites recovered was heterogeneous; certain parasite morphologies that have not been described previously were observed. In conclusion, the use of intraperitoneal diffusion chambers offers a potential tool for investigating thein vivodifferentiation process of secondary cysts ofEchinococcus granulosusin mice and the early local interactions between host and parasite during this process. 相似文献
995.
Junker BH Seamans TC Ramasubramanyan K Aunins J Paul E Buckland BC 《Biotechnology and bioengineering》1994,44(11):1315-1324
The titanium static mixer reactor, demonstrated for a variety of vaccine processes during the late 197s, was investigated for the production of attenuated hepatitis A virus antigen from anchorage-dependent MRC-5 cells. This reactor system used Charles River Biotechnological Services cabinets for monitoring and process control. Cell inoculation protocols, using 6000-10,000 cells/cm(2), resulted on over 95% attachment at both the laboratory and pilot scales. Indirect monitoring techniques using oxygen, glucose, L-serine, and L-glutamine uptake rates were indicative of cell growth prior to virus inoculation as well as environmental and/or nutrient limitations. Seven laboratory-scale (3900 cm(2)) runs and one pilotscale (265,000 cm(2)) run were conducted to investigate refeeding regiments, parallel versus perpendicular element orientation, increased element surface area per unit volume, and scale-up performance. In general, lysate antigen yields achieved were similar to those of parallel T-flasks cultivated under similar conditions. (c) 1994 John Wiley & Sons, Inc. 相似文献
996.
Fed-batch cultivation of animal cells using different medium design concepts and feeding strategies 总被引:4,自引:0,他引:4
In animal cell cultivation, cell density and product concentration are often low due to the accumulation of toxic end-products such as ammonia and lactate and/or the depletion of essential nutrients. A hybridoma cell line (CRL-1606) was cultivated in T-flasks using a newly devised medium feeding strategy. The goals were to decrease ammonia and lactate formation by the design of an initial medium which would provide a starting environment to achieve optimal cell growth. This was followed by using a stoichiometric equation governing animal cell growth and then designing a supplemental medium for feeding strategy used to control the nutritional environment. The relationship between the stoichiometric demands for glutamine and nonessential amino acids was also studied. Through stoichiometric feeding, nutrient concentrations were controlled reasonably well. Consequently, the specific production rate of lactate was decreased by fourfold compared with conventional fed-batch culture and by 26-fold compared with conventional batch culture. The specific production rate of ammonia was decreased by tenfold compared with conventional fed-batch culture and by 50-fold compared with conventional batch culture. Most importantly, total cell density and monoclonal antibody concentration were increased by five- and tenfold respectively, compared with conventional batch culture. (c) 1994 John Wiley & Sons, Inc. 相似文献
997.
On-line monitoring of monoclonal antibodies in animal cell culture using a grating coupler 总被引:1,自引:0,他引:1
Polzius R Bier FF Bilitewski U Jäger V Schmid RD 《Biotechnology and bioengineering》1993,42(11):1287-1292
A grating coupler was used for the on-line determination of monoclonal antibodies produced in perfused animal cell bioreactor. The device was connected with the culture vessel via a flow-injection analysis (FIA) system, which was controlled automatically. Specific antimouse lgG antibodies were immobilized on the surface of the sensor-chip. After injection of the sample, the binding of mouse lgG was observed in real time. The regeneration of the binding sites of the immobilized antibodies using an acidic solution allowed the on-line detection of produced monoclonal antibodies in the range of 10 to 150 mug/mL. In contrast to other techniques coupled to bioprocesses, the developed method represents a regenerable direct immunosensor. Results were compared with standard ELISA techniques (off-line) and a competitive immunochemical assay using the grating coupler (off-line). (c) 1993 John Wiley & Sons, Inc. 相似文献
998.
生殖抑制指原本具有生育能力的动物个体因特定外界环境或生理条件而减少或丧失生殖能力的现象,有时是受环境变化的主动调控,更多的是出现在其他个体影响下的被动抑制,极端情况发生在社会性动物的永久性抑制,即永久无法生殖或无法生殖成熟。研究发现非社会性动物也有生殖的推迟及可恢复性的生殖抑制,生殖抑制影响着动物种群数量动态、维持和进化。随着多学科的发展,生殖抑制机理研究取得了诸多进展。从阐述生殖抑制的概念出发,解析生殖抑制的形态、激素和分子生理特征,总结了生殖抑制的原因、作用,终述现有的理论模型以及不同物种方面的最新进展,并就生殖抑制在生物资源管理方面的应用价值进行了展望,旨在丰富生殖抑制的理论,扩展应用实践,为后续的生物资源管理提供理论参考。 相似文献
999.
Virginia Morera-Pujol Paulo Catry Maria Magalhães Clara Péron José Manuel Reyes-González José Pedro Granadeiro Teresa Militão Maria P. Dias Daniel Oro Giacomo Dell'Omo Martina Müller Vitor H. Paiva Benjamin Metzger Verónica Neves Joan Navarro Georgios Karris Stavros Xirouchakis Jacopo G. Cecere Antonio Zamora-López Manuela G. Forero Ridha Ouni Mohamed Salah Romdhane Fernanda De Felipe Zuzana Zajková Marta Cruz-Flores David Grémillet Jacob González-Solís Raül Ramos 《Diversity & distributions》2023,29(1):19-38
1000.
Joan-Beth Gow Shabnam Lainwala Timothy A. Lyerla 《In vitro cellular & developmental biology. Animal》1993,29(11):884-891
Summary Fibroblasts from a beige mouse (C57BL/6J;bg
J bgJ) have been established and maintained in culture for more than 3 yr. At early passages, the mutant cells were distinguishable
from C57BL/6J control mouse fibroblasts at the ultrastructural level by the presence of enlarged cytoplasmic granules. After
continuous passaging, this distinguishing feature was lost from the mutant cells, correlated with their increased growth rate.
Clustered, perinuclear distribution of lysosomes was retained, however, and was quantitatively different at any passage number
of the beige cell line from the dispersed distribution of these organelles in control mouse fibroblasts, as analyzed by computer-aided,
video-enhanced light microscopy. In somatic cell hybrids between the established beige cell line and a control human diploid
fibroblast cell strain, seven uncorrected hybrid lines retained a lysosomal dispersion pattern statistically indistinguishable
from that of the beige mouse cell lines. Three corrected hybrid lines had lysosomal dispersion patterns that were significantly
different from the beige parent line and indistinguishable from that of the control mouse fibroblast line. Thus, lysosomal
dispersion can be used objectively and quantitatively to distinguish mutant beige and control mouse fibroblasts and corrected
vs. uncorrected cell hybrids made from the beige/control human somatic cell crosses. 相似文献