Presents the latest advances in the biology and design of tissues and organs, while simultaneously connecting the basic sciences with the potential application of tissue engineering to diseases ...affecting specific organ systems.
The 19th ESACT meeting was to highlight the novel capabilities of the industry to move the products towards the clinic. It was attended by a wide range of workers in the industry and for many it was ...their first ESACT meeting. The proceedings here include the short papers adding the knowledge of the previous meetings and provide a reference for the researcher entering, or continuing in the field of Animal Cell Technology.
The demand for biopharmaceutical products is set to see a significant increase over the next few years. As a consequence, the processes used to produce these products must be able to meet market ...requirements. The present paper reviews the current technologies available for animal cell culture and highlights the advantages and disadvantages of each method, while also providing details of recent case studies. Processes are described for both suspension and anchorage‐dependent cell lines.
This works offers a simplified embodiment of the Stationary liquid mass balance method for the continuous estimate of the OUR estimation by means of the use of inexpensive proportional valves and the ...monitoring of their control signals is introduced and compared with the Global mass balance and Dynamic methods. As far said electrovalves can be considered to be linear, it can be demonstrated through the Mean value theorem for integrals that the mass balance equation for the liquid phase can be expressed as a first order differential equation: d C L dt = k L sm middot a sm middot ( α sm middot C L * - C L ) - OUR (i) Where: CL mol/l: DO concentration in the liquid phase C*L mol/l: DO concentration in equilibrium with the gas phase kL·a 1/h: Mass transfer coefficient a : PWM control signal or duty cycle Now, considering the DO concentration control loop error signal for a given constant set point. The previous equation (i) can be rewritten as a function of the control loop parameters and a new expression of the OUR proportional to the duty cycle and the mass transfer coefficient can be proposed: de dt = dC L sp dt - dCL dt C L sp = ctn implies de dt = dCL dt (ii) Where: emol/l: DO control loop error signal. CspLmol/l: DO set point OUR = k L sm middot a sm middot ( α sm middot C L * - C L ( t ) ) - de dt (iii)
Background: Altering target cell apoptosis is one of the challenging ideas of biotechnological applications. There are several applications of over expressing Bcl-xL anti-apoptotic protein from ...recombinant protein production to DNA vaccination strategies. The aim of the present study is to evaluate the anti-apoptotic efficacy of Bcl-xL expressing dual promoter plasmid system as a candidate to be used for recombinant protein production and DNA vaccination approaches. For this purpose, Bcl-xL anti-apoptotic protein gene was inserted in a dual expressing vector system in frame with EGFP (enhanced green fluorescence protein) after IRES (internal ribosomal site). The plasmid has a multiple cloning site after CMV (cytomegalovirus promoter) left empty to be inserted a biopharmaceutical protein gene region or DNA vaccine antigens. Results: In order to determine the anti-apoptotic efficacy of Bcl-xL inserted dual expressing vector, BHK-21 cells were transfected both with this plasmid and empty vector as control. Apoptosis was stimulated by several apoptosis inducing agents and serum deprivation in the transfected cells for 48 hrs. Cells expressing Bcl-xL protein in frame with EGFP were determined by flow cytometry as an indicator of cell viability. Additionally, apoptosis were determined by intracellular cleaved Casp 3 staining in Bcl-xL expressing EGFP positive cells. The dual expression plasmid bearing Bcl-xL anti-apoptotic protein prolonged the cell survival rate and protected cells from apoptosis upon apoptosis induction by doxorubicin and camptothecin in which the anti-apoptotic efficacies are inhibited through over expressing of Bcl-xL. pIRES2EGFP/Bcl-xL transfected cell ratio was significantly higher compared to empty vector transfected cells (P < 0.001). In contrast, apoptotic cell ratio was significantly lower in pIRES2EGFP/Bcl-xL transfected cell population compared to empty vector transfected cells (P < 0.001). Conclusion: In conclusion, it was shown that in vitro transient expression of Bcl-xL efficiently inhibited apoptosis induced by serum deprivation, doxorubicin and camptothecin. Thus, the dual expression plasmid bearing Bcl-xL anti-apoptotic protein could be a good candidate for recombinant protein production and DNA vaccination applications.
Animal cell technology is a growing discipline of cell biology which aims not only to understand structures, functions and behaviors of differentiated animal cells, but also to ascertain their ...abilities to be used for industrial and medical purposes. The goal of animal cell technology includes the clonal expansion of differentiated cells, the optimization of their culture conditions, modulation of their ability to produce proteins of medical and pharmaceutical importantance, and the application of animal cells to gene therapy, artificial organs and the production of functional foods. This volume gives the readers a complete review of the present state-of-the-art and will be useful for those working in either academic environments or in the biotechnology and pharmaceutical sectors, particularly cell biologists, biochemists, molecular biologists, immunologists, biochemical engineers and all other disciplines related to animal cell culture.
Cell Line Development Al-Rubeai, Mohamed
2009, 20090719, 2014-07-30, Letnik:
6
eBook
Mammalian cell lines command an effective monopoly for the production of therapeutic proteins that require post-translational modifications. This unique advantage outweighs the costs associated with ...mammalian cell culture, which are far grater in terms of development time and manufacturing when compared to microbial culture. The development of cell lines has undergone several advances over the years, essentially to meet the requirement to cut the time and costs associated with using such a complex hosts as production platforms. This book provides a comprehensive guide to the methodology involved in the development of cell lines and the cell engineering approach that can be employed to enhance productivity, improve cell function, glycosylation and secretion and control apoptosis. It presents an overall picture of the current topics central to expression engineering including such topics as epigenetics and the use of technologies to overcome positional dependent inactivation, the use of promoter and enhancer sequences for expression of various transgenes, site directed engineering of defined chromosomal sites, and examination of the role of eukaryotic nucleus as the controller of expression of genes that are introduced for production of a desired product. It includes a review of selection methods for high producers and an application developed by a major biopharmaceutical industry to expedite the cell line development process. The potential of cell engineering approch to enhance cell lines through the manipulation of single genes that play important roles in key metabolic and regulatory pathways is also explored throughout.
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