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Methods in molecular biology, vol. 5: Animal cell cultures
TIBTECH- DECEMBER 1990 [Vol. 8]
371
No solution to the Greenhouse Effect is possible until we change these wasteful policies. The changes could generate wealth, particularly for developing countries, but will require a major investment in research and development for biomass conversion. This book is an excellent starting point for that necessary effort. It reviews every relevant aspect of the topic from the volume, structure and composition of the crops themselves, through the biochemistry and microbiology of the fermentations, to the chemical engineering and []
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economics of currently practised processes. It contains a mass of facts and figures that should make it an essential sourcebook for libraries that claim to cover this important but under-studied field. Added to these are clear explanations, scholarly discussions and abundant references to relevant reviews in each of the many disciplines involved. I believe that this book will be widely useful, both as a source of reference and as a teaching aid in this field. It is scholarly; biotechnologists of all descriptions will find it useful, and so will chemical engineers. It is []
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Animal cell culture: a valuable working text M E T H O D S IN M O L E C U L A R B I O L O G Y , V O L . 5: A N I M A L CELL C U L T U R E S
edited by Jeffrey W. Pollard and John M. Walker, Humane Press, 1989. $69.50 (xiv + 713 pages) ISBN 0 89603 150 0 This volume in the series 'Methods in Molecular Biology' provides a comprehensive collection of state-ofthe-art techniques for the culture of various animal cell types. Most chapters not only cover the basic methodology but also include highly specialist techniques and conditions for cell culture in vitro. Clear, detailed protocols (including sources of reagents) are set out throughout the volume. Procedures for establishing extracellular matrices to maintain the differentiated functions of cultured cells should have wide applications for the culture of 'fastidious' cell types. Agar-culture systems for the clonal growth and differentiation of haemopoietic cells, the establishment of long-term bone marrow cultures and growth-factordependent haemopoietic cells are clearly described. These culture systems provide valuable models for fundamental research in haemopoiesis. Other important techniques described include cine microscopy, electron microscopy, immunolabelling, cytogenetics, flow cytometry and fluorescence-activated cell sorting. A useful and interesting technique for significantly improving transfection efficiencies in human thy-
roid epithelial cells is described in Chapter 42. The trick is to add thyroid-stimulating hormone (TSH) when transfecting DNA by co-precipitation with calcium (or strontium) phosphate. Whilst calcium phosphate co-precipitation is an established transfection technique; it is by no means a trivial exercise; buffer pH, DNA quality and concentration, incubation time and cellular toxicity to calcium are all critical parameters. These need to be carefully titrated for optimal transfection efficiency. The information in this chapter is clear but a comparison of this technique with cationic liposome mediated transfection would have been useful. In addition, we would have liked to know if the TSH manoeuvre works with other cell types. There is a useful introductory chapter on cloning foreign genes using the baculovirus expression system in insect cells. The authors use pAc373 as their example cloning vector and pAc360 as an example of a polyhedrin gene fusion vector. Mention could have been made of some of the other baculovirus vectors now available. These employ viral promoters that are active at different stages of the insect cell lifecycle. These vectors could be usefully employed if there are problems with levels of expression, with solubility, or doubts about post-translational modification. In addition, vectors that enable the expression of more than one gene are now available and in use. Cationic liposome sys-
not polemic and only mildly visionary, but policy makers, industrialists and economists should try to read it, because it gives facts about volume, price and prospects for products that they will find it difficult to amass from elsewhere. In short, it is a background for discussion of the development of the future biomass industry that will eventually arise. B. S. H A R T L E Y
Centre for Biotechnology, Imperial College of Science and Technology, South Kensington, London SW7 2AZ, UK. []
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tems (e.g. Lipofectin) instead of calcium co-precipitation is another alternative and, in many ways, a preferable method of transfection in this system. The chapters covering hybridomas, their cryopreservation screening and the isotyping of monoclonal antibodies are very clear and concise. An ELISA system is outlined as a means for screening antibodies in hybridoma supernatants. An accompanying protocol and diagram are clear. Other topics c o v e r e d include monoclonal antibody purification and an introduction to hybridoma production in rats. What is possibly missing from this section of the book is a chapter covering anti-peptide antibodies, their usefulness, construction, production and purification. This is an extremely powerful technique for obtaining antibodies against proteins when the native protein antigen is not available. As a whole, the volume is a valuable working text for researchers in the general area of animal cell culture. The protocols are particularly clear and are supported with diagrams, pictures and references. It will be useful to both new workers and those more experienced in this field. D. K N I G H T G. S H A H G. G O U G H
Departments of Natural Products and Virology, Glaxo Research Group, Greenford Road, Greenford, Middlesex UB6 0HE, UK.
371
No solution to the Greenhouse Effect is possible until we change these wasteful policies. The changes could generate wealth, particularly for developing countries, but will require a major investment in research and development for biomass conversion. This book is an excellent starting point for that necessary effort. It reviews every relevant aspect of the topic from the volume, structure and composition of the crops themselves, through the biochemistry and microbiology of the fermentations, to the chemical engineering and []
[]
[]
economics of currently practised processes. It contains a mass of facts and figures that should make it an essential sourcebook for libraries that claim to cover this important but under-studied field. Added to these are clear explanations, scholarly discussions and abundant references to relevant reviews in each of the many disciplines involved. I believe that this book will be widely useful, both as a source of reference and as a teaching aid in this field. It is scholarly; biotechnologists of all descriptions will find it useful, and so will chemical engineers. It is []
[]
[]
[]
[]
[]
Animal cell culture: a valuable working text M E T H O D S IN M O L E C U L A R B I O L O G Y , V O L . 5: A N I M A L CELL C U L T U R E S
edited by Jeffrey W. Pollard and John M. Walker, Humane Press, 1989. $69.50 (xiv + 713 pages) ISBN 0 89603 150 0 This volume in the series 'Methods in Molecular Biology' provides a comprehensive collection of state-ofthe-art techniques for the culture of various animal cell types. Most chapters not only cover the basic methodology but also include highly specialist techniques and conditions for cell culture in vitro. Clear, detailed protocols (including sources of reagents) are set out throughout the volume. Procedures for establishing extracellular matrices to maintain the differentiated functions of cultured cells should have wide applications for the culture of 'fastidious' cell types. Agar-culture systems for the clonal growth and differentiation of haemopoietic cells, the establishment of long-term bone marrow cultures and growth-factordependent haemopoietic cells are clearly described. These culture systems provide valuable models for fundamental research in haemopoiesis. Other important techniques described include cine microscopy, electron microscopy, immunolabelling, cytogenetics, flow cytometry and fluorescence-activated cell sorting. A useful and interesting technique for significantly improving transfection efficiencies in human thy-
roid epithelial cells is described in Chapter 42. The trick is to add thyroid-stimulating hormone (TSH) when transfecting DNA by co-precipitation with calcium (or strontium) phosphate. Whilst calcium phosphate co-precipitation is an established transfection technique; it is by no means a trivial exercise; buffer pH, DNA quality and concentration, incubation time and cellular toxicity to calcium are all critical parameters. These need to be carefully titrated for optimal transfection efficiency. The information in this chapter is clear but a comparison of this technique with cationic liposome mediated transfection would have been useful. In addition, we would have liked to know if the TSH manoeuvre works with other cell types. There is a useful introductory chapter on cloning foreign genes using the baculovirus expression system in insect cells. The authors use pAc373 as their example cloning vector and pAc360 as an example of a polyhedrin gene fusion vector. Mention could have been made of some of the other baculovirus vectors now available. These employ viral promoters that are active at different stages of the insect cell lifecycle. These vectors could be usefully employed if there are problems with levels of expression, with solubility, or doubts about post-translational modification. In addition, vectors that enable the expression of more than one gene are now available and in use. Cationic liposome sys-
not polemic and only mildly visionary, but policy makers, industrialists and economists should try to read it, because it gives facts about volume, price and prospects for products that they will find it difficult to amass from elsewhere. In short, it is a background for discussion of the development of the future biomass industry that will eventually arise. B. S. H A R T L E Y
Centre for Biotechnology, Imperial College of Science and Technology, South Kensington, London SW7 2AZ, UK. []
[]
[]
tems (e.g. Lipofectin) instead of calcium co-precipitation is another alternative and, in many ways, a preferable method of transfection in this system. The chapters covering hybridomas, their cryopreservation screening and the isotyping of monoclonal antibodies are very clear and concise. An ELISA system is outlined as a means for screening antibodies in hybridoma supernatants. An accompanying protocol and diagram are clear. Other topics c o v e r e d include monoclonal antibody purification and an introduction to hybridoma production in rats. What is possibly missing from this section of the book is a chapter covering anti-peptide antibodies, their usefulness, construction, production and purification. This is an extremely powerful technique for obtaining antibodies against proteins when the native protein antigen is not available. As a whole, the volume is a valuable working text for researchers in the general area of animal cell culture. The protocols are particularly clear and are supported with diagrams, pictures and references. It will be useful to both new workers and those more experienced in this field. D. K N I G H T G. S H A H G. G O U G H
Departments of Natural Products and Virology, Glaxo Research Group, Greenford Road, Greenford, Middlesex UB6 0HE, UK.