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Author : Maureen A. Harrison,Ian F. Rae
Publisher : Cambridge University Press
Page : 178 pages
File Size : 53,5 Mb
Release : 1997-10-13
Category : Science
ISBN : 052157496X
Concise introduction to a major technique of cell biology laboratories for those new to the field.
Author : Bernice M. Martin
Publisher : Springer Science & Business Media
Page : 251 pages
File Size : 43,6 Mb
Release : 2013-12-01
Category : Medical
ISBN : 9781461202479
ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Xl II INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I STERILITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Aseptic Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Physical manipulations • Use of the sterile cabinet (hood) Sterilization Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Heat • Radiation • Toxic gas • Filtration • Antibiotics Quality Control of Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 Routine labeling Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 vi CONTENTS ROUTINE CELL CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Feeding Schedules and Media Components . . . . . . . . . . . . . . . . . . . . 29 General properties of media and salt solutions • water as a reagent· Establishingfeeding schedules Subcultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Solutions and methods for adherent cells • Common enzyme solutions • Inoculating (seeding) the cultures Cell Enumeration and Cell Viability . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Hemocytometer • Particle counter • Cell viability Putting Routine Methods to Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Normal cell growth characteristics Detecting and Disposing of Contamination . . . . . . . . . . . . . . . . . . . . 66 Bacteria and fUngi • Fungi • Mycoplasma • Viruses • Dealing with contamination Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 Inadequate cell growth • Recurrent contamination • When to call your vendor Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Biological hazards • Chemical hazards Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 EXPERIMENTS IN CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . 91 II Alterations of the Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Serum • Treatments of serum • Plasma-derived serum • Serum-free and low-protein media Substrata. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 Coatingplasticware with solutions • Alterations with polymers • Using cells to coat the plasticware • Culturing cells on microcarriers Altering the Environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Temperature changes • Gaseous changes Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 CONTENTS vii PRIMARY CELL CULTURE. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Dissection • Enzymatic dissociation methods • Nonenzymatic isolation • Purification of cell suspensions • Consideringyield and survival Chatacterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Author : Michael R. Davey,Paul Anthony
Publisher : John Wiley & Sons
Page : 358 pages
File Size : 50,8 Mb
Release : 2010-05-20
Category : Science
ISBN : 0470686510
The ability to culture cells is fundamental for mass propagation and as a baseline for the genetic manipulation of plant nuclei and organelles. The introduction to Plant Cell Culture: Essential Methods provides a general background to plant cell culture, including basic principles, technologies and laboratory practices that underpin the more detailed techniques described in subsequent chapters. Whilst each chapter provides a background to the topic area and methodology, a crucial aspect is the provision of detailed protocols with emphasis on trouble shooting, describing common problems and detailed advice for their avoidance. Plant Cell Culture: Essential Methods provides the reader with a concise overview of these techniques, including micropropagation, mutagenesis, cryopreservation, genetic and plastid transformation and somatic cell technologies. This book will be an essential addition to any plant science laboratory's bookshelf. Highlights the best and most up-to-date techniques for working on plant cell culture Explains clearly and precisely how to carry out selected techniques in addition to background information on the various approaches Chapters are written by leading international authorities in the field and cover both well-known and new, tried and tested, methods for working in plant cell culture An essential laboratory manual for students and early-career researchers.
Author : Martin Clynes
Publisher : Springer Science & Business Media
Page : 624 pages
File Size : 43,6 Mb
Release : 2012-12-06
Category : Science
ISBN : 9783642804120
Cell culture techniques allow a variety of molecular and cell biological questions to be addressed, offering physiological conditions whilst avoiding the use of laboratory animals. In addition to basic techniques, a wide range of specialised practical protocols covering the following areas are included: cell proliferation and death, in-vitro models for cell differentiation, in-vitro models for toxicology and pharmacology, industrial application of animal cell culture, genetic manipulation and analysis of human and animal cells in culture.
Author : Jennie P. Mather,Penelope E. Roberts
Publisher : Springer Science & Business Media
Page : 247 pages
File Size : 54,6 Mb
Release : 2007-08-20
Category : Science
ISBN : 9780585275710
It is a pleasure to contribute the foreword to Introduction to Cell and Tissue Culture: The ory and Techniques by Mather and Roberts. Despite the occasional appearance of thought ful works devoted to elementary or advanced cell culture methodology, a place remains for a comprehensive and definitive volume that can be used to advantage by both the novice and the expert in the field. In this book, Mather and Roberts present the relevant method ology within a conceptual framework of cell biology, genetics, nutrition, endocrinology, and physiology that renders technical cell culture information in a comprehensive, logical for mat. This allows topics to be presented with an emphasis on troubleshooting problems from a basis of understanding the underlying theory. The material is presented in a way that is adaptable to student use in formal courses; it also should be functional when used on a daily basis by professional cell culturists in a- demia and industry. The volume includes references to relevant Internet sites and other use ful sources of information. In addition to the fundamentals, attention is also given to mod ern applications and approaches to cell culture derivation, medium formulation, culture scale-up, and biotechnology, presented by scientists who are pioneers in these areas. With this volume, it should be possible to establish and maintain a cell culture laboratory devot ed to any of the many disciplines to which cell culture methodology is applicable.
Author : Simon P. Langdon
Publisher : Springer Science & Business Media
Page : 346 pages
File Size : 53,9 Mb
Release : 2008-02-01
Category : Medical
ISBN : 9781592594061
The culture of cancer cells is routinely practiced in many academic research centers, biotechnology companies, and hospital laboratories. Cancer Cell Culture: Methods and Protocols describes easy-to-follow methods to guide both novice and more experienced researchers seeking to use new techniques in their laboratories. Our present understanding of the cell and molecular biology of cancer has been derived mainly from the use of cultured cancer cells and we cover a number of the most widely used assays to study function in current use. Part I introduces the basic concept of cancer cell culture and this is followed by a description of the general techniques used in many cell culture facilities. The importance of cell line characterization is now widely recognized and methods to characterize and authenticate cell lines are described in Part II. Part III covers the isolation and development of specific cancer cell types and provides valuable tips for those wishing to derive new cell line models. A wide range of procedures encompassing many of the key functional features of cancer cells are described in Part IV including assays to evaluate clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters. Methods to modify cancer cells are described in Part V, including protocols for transfection, development of drug-resistance, immortalization, and transfer in vivo. In Part VI methods of coculture of different cell types and contamination of cell lines are covered.
Author : Víctor M. Loyola-Vargas,Felipe Vázquez-Flota
Publisher : Springer Science & Business Media
Page : 389 pages
File Size : 53,9 Mb
Release : 2008-02-04
Category : Science
ISBN : 9781592599592
A comprehensive state-of-the-art collection of the most frequently used techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. Two new appendices explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, and list more than 100 very useful internet sites.
Author : Jun Mitsuhashi
Publisher : Springer Science & Business Media
Page : 435 pages
File Size : 53,6 Mb
Release : 2012-12-06
Category : Science
ISBN : 9784431678755
I started insect cell culture work in 1962, when T. D. C. Grace reported the first establishment of invertebrate continuous cell lines. He obtained grow ing cells from pupal ovaries of the emperor gum moth, Antheraea euca lypti. At that time, I was trying to obtain growing cells from leafhoppers. Grace's method could not be applied directly to my culture because of the differences in species, the size of the insects, and the tissue to be cul tured. The vertebrate tissue culture methods gave me some ideas for pre paring cultures from leafhoppers, but those could not be used directly either. There were no textbooks and no manuals for invertebrate tissue culture, so I had to develop a method by myself. First, I considered what type and what size of vessels are suitable for insect tissue culture. Also, I had to look for suitable materials to construct the culture vessels. Sec ond, I had to examine various culture media, especially growth-promot ing substances, such as sera. Then I had to improve culture media by trial and error. The procedure to set up a primary culture was also a problem. How could I sterilize materials? How could I remove tissues from a tiny insect? How many tissues should I pool in order to set up one culture? I had to find out the answers. Naturally, it took a lot of time.
Author : E. N. Willmer
Publisher : Elsevier
Page : 804 pages
File Size : 47,8 Mb
Release : 2015-12-04
Category : Science
ISBN : 9781483223001
Cells and Tissues in Culture: Methods, Biology and Physiology, Volume 1 covers the general fields of tissue culture, including an evaluation of its technique, effects, and contributions to biology. This book focuses on the three methods of culture—tissue culture, cell culture, and organ culture.Other topics include the design of complete synthetic media, possible evolution of the cell types, and energy relationships in growing and stationary cells. The RNA synthesis in cell cultures, culture of amphibian embryonic anlage, action of corticosteroids and adrenaline, and effects of parathyroid hormone on bone are also elaborated. This volume is recommended for biologists and specialists interested in the culture of cells and tissues.
Author : Shohreh Amini,Martyn K. White
Publisher : Humana
Page : 205 pages
File Size : 42,7 Mb
Release : 2022-06-09
Category : Medical
ISBN : 1071614398
This second edition volume details the latest aspects of neural cells covering the practical and theoretical considerations of each techniques involved. Chapters guide readers through a general overview of the neuronal culturing principles, cell line models for neural cells, the isolation and propagation of primary cultures, stem cells, transfection and transduction of neural cultures, and other more advanced techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and easy to use, Neuronal Cell Culture: Methods and Protocols, Second Edition aims to be of interest to scientists at all levels studying cell culture models for neuroscientific studies.
Author : Donald Joseph Merchant
Publisher : Unknown
Page : 212 pages
File Size : 52,6 Mb
Release : 1960
Category : Cultures (Biology)
ISBN : WISC:89041307018
General laboratory intructions; Cell culture techniques; Organ culture methods; Applications of cell and organ culture; Techniques and procedures; Media and reagents.
Author : Bruce Alberts
Publisher : Unknown
Page : 0 pages
File Size : 49,8 Mb
Release : 2002
Category : Cytology
ISBN : 0815332181
Author : Paul F. Jr. Kruse
Publisher : Elsevier
Page : 897 pages
File Size : 47,5 Mb
Release : 2012-12-02
Category : Science
ISBN : 9780323142076
Tissue Culture: Methods and Applications presents an overview of the procedures for working with cells in culture and for using them in a wide variety of scientific disciplines. The book discusses primary tissue dissociation; the preparation of primary cultures; cell harvesting; and replicate culture methods. The text also describes protocols on single cell isolations and cloning; perfusion and mass culture techniques; cell propagation on miscellaneous culture supports; and the evaluation of culture dynamics. The recent techniques facilitating microscopic observation of cells; cell hybridization; and virus propagation and assay are also encompassed. The book further tackles the production of hormones and intercellular substances; the diagnosis and understanding of disease; as well as quality control measures. Scientists and professionals interested in methodology per se will find the book invaluable.
Author : Glyn Stacey,Alan Doyle,Margherita Ferro
Publisher : Springer Science & Business Media
Page : 166 pages
File Size : 43,6 Mb
Release : 2001-12-31
Category : Medical
ISBN : 1402004273
Cell Culture Methods for in vitro Toxicology introduces the reader to a range of techniques involved in the use of in vitro cell culture in toxicological studies. It deals with major cell types studied in the field of toxicology and will be useful for anyone wishing to start work with animal cell cultures or to refresh their knowledge relating to in vitro cell models. Fundamental chapters deal with the general biology of cytotoxicity and cell immortalisation these are key issues for in vitro systems addressing the `3Rs' principle. Up-to-date overviews deal with the use of cells from liver, brain and intestine. In addition, biochemical analysis of cell responses, biotransformation pathways in cells and recombinant approaches to the early detection of cell stress are also covered in detail. Prominent features of in vitro technologies also include regulation, biosafety and standardisation. Dedicated chapters deal with these issues in a practical way in order to lead the reader to the right source of information. This book provides an up-to-date, informative and practical review of cell culture methods for in vitro toxicology. It will be of equal benefit to students and experienced toxicologists with little experience of in vitro cell culture.
Author : R. Ian Freshney
Publisher : Wiley-Liss
Page : 510 pages
File Size : 55,7 Mb
Release : 1993-12-29
Category : Science
ISBN : 0471589667
This masterful third edition of Freshney's Culture of Animal Cells updates and considerably expands the scope of its predecessor and still enables both the novice and the experiences researcher to apply the basic and more sophisticated techniques of tissue culture. New Topics covered include: the use of molecular techniques in cell culture, such as DNA fingerprinting, fluorescence in situ hybridization, and chromosome painting cell interactions in cell culture new methods for separating cells new or refined methods for accessing cytotoxicity, viability, and mutagenicity experimental details for culture of specialized cells types not covered in previous editions new or refined techniques for visualizing clues, including time-lapse photography and confocal microscopy The revised and expanded third edition offers the following features: over 350 new reference to the primary literature an international list of cell banks an international listing of reagants and commercial supplies a subject index a glossary Also available: 0471169021 Culture of Animal Cells: A Multimedia Guide CD-ROM $150 est. From the reviews: "I strongly recommend this volume for any laboratory wishing to culture mammalian cells" - Biotechnology "It is not very often that it is possible to say of a book, 'I don't know how I managed without it previously.' Here is such a book" - Cell Biology International Reports
