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Techniques for long life Culture of hmnortalised Cells

edited by R. Ia, Fresh,e), and Mary (;. Freshne),,John ll;)ley & So~s, 1996.

UKJ]40. O0 (389 pases) ISBN 0 471 1213 47

Cells that are cultured from uonnal somatic tissue do not live for ever: individual cells undergo a set number of divisions (the Hayflick limit) before they senesce and die. By contrast, cells that are cultured from malignant tunmurs do not have a limited lifespan and, given the right culture conditions, will grow indefinitely. Sometimes nomlal cells will undergo a malignant transformation iu Ntro and, when this happens, they lose the phenotypic characteristics of normal cells and develop a malignant phenotype with unlimited growth. At one time it was assumed that transformation to a malignant phenotype was essential for the development of immortalized cell lines. Recently, however, techniques have been developed that allow normal cells to become immortalized in culture whilst retaining at least some of the 'normal' phenotypic characteristics. Such cells are invaluable in biotechnology, not least because they can carry out the post-translational modifications

to genetically engineered proteins which are unavailable in prokaryotic systems. hnmortalization of normal cells is achieved by infecting cells with viruses, such as simian virus 40 (SV40) or Epstein-Barr virus (EBV) (the latter for immortalizing B cells), or by transf~-cting the cells with innnortalizing genes, such as the SV40 large T antigen gene.

Cultme of lmmortalised Cells is the third book in a series that deals with the culture of specialized cells. It is a truly international book with contributions from specialists around the world. The book begins with a somewhat complex review of the current thinking on the genetic basis for cellular senescence aim imnlortality, the latter particularly in relation to the immortalization of keratinocytes. The bulk of the book (13 chapters) contains protocols for producing immortalized cells from a varie D' of (mostly human) sources including hepatocytes, astrocytes, keratinocytes and glial cells. A further chapter details methods

available for mapping human senescence genes. Each of the 'methods' chapters contains useful introductory material, though the amount varies considerably. The protocols themselves are presented in a clear and straightforward manner, which should be relatively easy to follow, and there is surprisingly little overlap between chapters. Each chapter is fully referenced, though it is claimed that the detailed protocols obviate the need for primary" literature sources. In addition, there is a very useful and timely chapter on safety" aspects of working with immortalized cells.

Cuhure of lmmortalised Cells assumes a good working knowledge of tissue culture and molecular biological techniques and therefore cannot really be aimed at undergraduates. Given the nature of the work and the high level at which it is written, it will be invaluable for research personnel in tissue culture laboratories where cell immortalization is being contemplated. O f particular value is the variety- of cell sources covered, allowing a good overview of what has been achieved so far, and the comprehensive infomaation on methods, reagents and suppliers. In summary, an excellent source of infomlation in a highly specialized field.

Maureen Dawson Department of Biological Sciences,

Manchester Metropolitan University, Manchester, UK M1 5GD.

Organotypic protocols Epithelial Cell Culture: A Practical Approach

edited by A. J. Shau,, Oxfi~rd ( hliversity Press, 1996. U K ~ 2 7.50 (238 pq~es)

ISBN 0 19 96 3572 2 (paperback) (7K:£55.00 ISBN 0 19 963573 0

(hardback)

Call me old-fashioned but I alveays like to start reading a book at the beginning, with the preface. In this case this was a veW good place to start as Dr Shaw's aims for the book are spelled out quite concisely:

• to provide detailed protocols for modellin S [my emphasis] different epithelia in vitro using primary cultures or cell lines; • to provide detailed protocols for morpholo~cally, biochemically and functionally characterizing these

models in relation to their in vivo counterparts; and • to outline research applications to which these cultures can be applied.

The clue to the real nature of this book comes earlier in the preface where Dr Shaw points out that a common theme is the culture of epithelia on porous substrates, so it should really be called: 'OrganotTpic Culture: A Practical Approach' - but then it would probably not sell well. This would be a shame because Dr Shaw has put together a fine collection of chapters that more than adequately meet his aims.

Chapter 1 serves as an excellent introduction to the nature of

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book re ,ieues

epkhelia (although I am sceptical about Dr Shaw's efforts to include encothelial cells under this banner), and, whilst it does not contain any protocols per se, does serve as a use~ul reference source for the coraing chapters. One item that might usefully have been included here would be a discussion of how prima W biopsy material, particularly human (although handling bovine brain for the protocols in Appendix 2 might presently be corLsidered hazardous too!), may be abtained and handled. There are ethical and safety, issues here that are not adequately addressed in anv of the remaining chapters. I also found it slightly disappointing that no reference to systems for culture of breast epithelium is made here (or indeed, anywhere else before page 136). These minor criticisms aside, Chapter 1 does whet the appetite for what follows, and what follows are eight chapters and appendices describing the culture, characterization and uses of human kmatinocytes, canine kidney epithelial cells, human and rat

hepatocytes, a human intestinal epithelial cell line, guinea pig endometrial cells, human endothelial cells, rat Sertoli cells and bovine brain microvascular endothelial cells. Each of these chapters contains the expected detailed and well illustrated protocols, each is well referenced and, with one exception, well introduced and discussed. The exception is Chapter 4 on xenobiotic metabolism, in which the authors launch into the protocols without adequately introducing the subject. Nowhere can the uninitiated find a description of the nature of Phase I and II reactions, or what they are important for, or why one should wish to assess them in hepatocytes.

It is clear that this volume is targeted at those scientists who wish to study toxicolog% pharmacology and pharmaceutics in l~itro. It will therefore appeal to those engaged in industrial applications as well as those engaged in more basic research. It is also clearly targeted predominantly at the young - who

else could decipher the tiny font used to list the ingredients at the top of each protocol?

Other reviewers of books in this series have carped at the lack of telephone or fhx numbers for the suppliers listed in the appendices. I am not convinced these would be useful as there are often many contact numbers and, in the UK at least, these are prone to change with monotonous regularity. However, it might be useful to include World Wide Web site addresses as a single point of reference in future.

My recommendation? If you are interested in organotypic cultures of epithelial cells, buy this book and it wiU serve you well. If you are interested in more basic epithelial cell cu]ture, others might be of more use.

Ian McKay Academic Department of Dermatology,

St Bartholomew's and the Royal London School of Medicine and Dentistry, Queen

Mary & Westfield College, London, UK E1 2AT.

Cutting proteins down to size

Industrial Enzymolog~ (2nd edn)

edited b), T. Godfre), and S. H~'st, Macmillan, 1996. UK£110 . O0

&il, + 609p,L~es) I S B N 1 56159 163 7

Since the first edition of this book appeared in 1983, there have been many significant developments in en::ymolog2y and related fields. The editors have, therefore, updated the previous edition and, in this volume, provide a comprehensive act ount of current practice in incustrial enzymolog% with a topical focus on environmental aspects, concerns with food safety,, and newer applications such as biosensor development. The book opens with a brief introduction to the subject, emphasizing commercial aspects such as current/projected markets for industnal enzymes and factors important in selecting enzymes for particular processes. The bulk of the book (almost 400

pages) focuses on detailed descriptions of applications of enzymes to some 22 industrial sectors, ranging from biosensors to textiles. It concludes with chapters on legislative, toxicological and safew aspects of the topic and with a description of applied enzyme kinetics. In addition, there is a lot of compiled information provided as tables or appendices, such as pH/temperature properties of industrial enzymes, names and addresses of commercial suppliers of enzymes and other products and a guide to the use of enzwnes m the more common industrial areas. This is a valuable addition to the literature of applied enzymoloD'.

In general, I found the treatment to be comprehensive and readable. The editors have done a fine job m producing a multi-author volume with a high deg-ree of internal consistency in terms of style and content. With the exception of a small number of annoying typog-raphical errors m parts of the book, it is produced to quite a high standard with frequent use of tables, drawings and chenrical structures. The book is not without its flaws, though, in the light of developments in protein expression systems and downstream processing of proteins in the last 13 years, I was surprised that no coverage was given to the actual industrial production of enzymes f~3r the industries described. This could have provided an important unifying strand through the book and would have given the opportuniw of exploring the vauing levels of purity/activity, required for different applications (for example in diagnostic application compared, say, to use in a food-related process). Several contributors allude to variation in properties of industrial

TIBTECH MARCH 1997 (VOL 15}