Human Cell Culture Protocols

Human cell culture is not a new topic, but the development of new molecular techniques and reagents which can be used to investigate cell function and the responsible intracellular mechanisms make it a continuing requirement. This third edition of Human Cell Culture Protocols expands upon the previous editions with current, detailed protocols for the isolation and culture of a range of primary cells from human tissues. With new chapters on pancreatic cells needed for basic studies on the pathogenesis of diabetes and for their application for islet transplantation, the book also delves into protocols for hepatocytes, skin cells, lung cells, parathyroid cells, gastric cells, renal cells, adipocytes, ovarian cells, bone cells, vascular smooth muscle cells, vascular endothelial cells, regulatory T cells, blood mononuclear cells, as well as new techniques being applied to human cell culture, particularly the use of biocompatible scaffolds to grow cells, the in vitro use of laser microdissection to isolate cells from culture, and automated cell culture. Written in the highly successful Methods in Molecular BiologyTM 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. Authoritative and cutting-edge, Human Cell Culture Protocols, Third Edition makes it possible for a worker with basic cell culture training, whether in the fields of cell biology, gene therapy, and cell transplantation, to prepare cell cultures of the specific cell type necessary to forward their vital research.

A thoroughly revised and updated collection readily reproducible techniques for culturing human cells. This new edition includes a wide range of human cell types relevant to human disease and new chapters on fibroblasts, Schwann cells, gastric and colonic epithelial cells, and parathyroid cells. The protocols follow the successful Methods in Molecular MedicineTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Now completely revised and updated from the original, much-acclaimed and bestselling first edition, Basic Cell Culture Protocols, 2nd ed. offers today's most comprehensive collection of easy-to-follow, cutting-edge protocols for the culture of a wide range of animal cells. Its authoritative contributors provide explicit, step-by-step instructions, along with extensive notes and tips that allow both experts and beginners to successfully achieve their desired results. Topics range from basic culture methodology to strategies for culturing previously uncultured cell types and hard-to-culture differentiated cells. Methods are also provided for the analysis of living cells by FACS, video microscopy, and confocal microscopy. Like the first edition, this book should be in every cell culture laboratory and be of use to all who use cell cultures in research.

At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. Written in the highly successful Methods in Molecular BiologyTM, 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. Comprehensive and up-to-date, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.

Fascinating biology occurs at epithelial interfaces, whether between organism and environment or within body compartments, and many diseases inflicting huge personal and societal burdens result from dysfunction of epithelial systems, e.g., carcinomas. Epithelial cell cultures have been an integral and crucial part of the biomedical research enterprise, adding unique capabilities and enabling mechanistic approaches. In the past decade there have been many research advances, such as directed differentiation of embryonic stem cells and induced pluripotent stem cells, robotic high throughput screening, whole genome siRNA and shRNA libraries, massively parallel sequencing at low cost, identification of somatic stem cells in key organs, to name a few. Epithelial Cell Culture Protocols, Second Edition provides a cross section of up-to-date culture protocols for the most heavily studied cell systems and featured supporting technologies. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Epithelial Cell Culture Protocols, Second Edition will serve outstanding investigators with the best possible information for the advancement of biomedical science.

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.

This manual is designed to serve as a practical guide to primary human cell culture, which is integral in both academic and industrial biotechnology research. As in the first edition, the content of the manual is not exhaustive, but rather contains selected protocols for specific cell types from major tissue groupings in the body. This improved second edition also includes a new section on stem cells and additional material on transfection. It should serve as a foundation for individual researchers to experiment, explore, and establish niche protocols for their specific needs. With its compact physical format that makes it portable and flexible for usage in a laboratory setting, the manual will be a useful guide for all beginners in primary human cell culture work.

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.

This manual is designed to serve as a practical guide to primary human cell culture, which is integral in both academic and industrial biotechnology research. As in the first edition, the content of the manual is not exhaustive, but rather contains selected protocols for specific cell types from major tissue groupings in the body. This improved second edition also includes a new section on stem cells and additional material on transfection. It should serve as a foundation for individual researchers to experiment, explore, and establish niche protocols for their specific needs. With its compact physical format that makes it portable and flexible for usage in a laboratory setting, the manual will be a useful guide for all beginners in primary human cell culture work.

With many recent advances, cancer cell culture research is more important than ever before. This timely edition of Cancer Cell Culture: Methods and Protocols covers the basic concepts of cancer cell biology and culture while expanding upon the recent shift in cell culture methods from the generation of new cell lines to the use of primary cells. There are methods to characterize and authenticate cell lines, to isolate and develop specific types of cancer cells, and to develop new cell line models. Functional assays are provided for the evaluation of clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters. Other methods permit the modification of cells for transfection, drug resistance, immortalization, and transfer in vivo, the co-culture of different cell types, and the detection and treatment of contamination. In this new edition, specific emphasis is placed on safe working practice for both cells and laboratory researchers. These chapters contain the information critical to success – only by good practice and quality control will the results of cancer cell culture improve. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Cancer Cell Culture: Methods and Protocols serves as a practical guide for scientists of all backgrounds and aims to convey the appropriate sense of fascination associated with this research field.