Animal Cell Electroporation and Electrofusion Protocols

1. Front Matter

   Pages i-xx

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2. Theory and Instrumentation

   1. Electroporation Theory

      • James C. Weaver

      Pages 3-28

   2. Effects of Pulse Length and Strength on Electroporation Efficiency

      • Sek Wen Hui

      Pages 29-40

   3. Instrumentation

      • Gunter A. Hofmann

      Pages 41-59

3. Electroporation Protocols

   1. The Introduction of Proteins into Mammalian Cells by Electroporation

      • William F. Morgan, Joseph P. Day

      Pages 63-71

   2. Electroporation of Antigen-Presenting Cells for T-Cell Recognition and Cytotoxic T-Lymphocyte Priming

      • Weisan Chen, James McCluskey

      Pages 73-81

   3. Electroporation of Antibodies into Mammalian Cells

      • Paul L. Campbell, James McCluskey, Jing Ping Yeo, Ban-Hock Toh

      Pages 83-92

   4. Electroporation of Adherent Cells In Situ for the Introduction of Nonpermeant Molecules

      • Leda H. Raptis, Kevin L. Firth, Heather L. Brownell, Andrea Todd, W. Craig Simon, Brian M. Bennett et al.

      Pages 93-113

   5. Electrotransformation of Chinese Hamster Ovary Cells

      • Danielle Gioioso Taghian, Jac A. Nickoloff

      Pages 115-121

   6. Electroporation of Rat Pituitary Cells

      • Ruth H. Paulssen, Eyvind J. Paulssen, Kaare M. Gautvik

      Pages 123-131

   7. Electroporation of Plasmid DNA into Normal Human Fibroblasts

      • F. Andrew Ray

      Pages 133-140

   8. Electroporation-Mediated Gene Transfer into Hepatocytes

      • Alphonse Le Cam

      Pages 141-150

   9. Electroporation of Human Lymphoblastoid Cells

      • Fen Xia, Howard L. Liber

      Pages 151-160

   10. The Use of Electroporated Bovine Spermatozoa to Transfer Foreign DNA into Oocytes

       • Marc Gagné, François Pothier, Marc-André Sirard

       Pages 161-166

   11. Electroporation of Embryonic Stem Cells for Generating Transgenic Mice and Studying In Vitro Differentiation

       • John S. Mudgett, Thomas J. Livelli

       Pages 167-184

   12. Electrotransfection with “Intracellular” Buffer

       • Maurice J. B. van den Hoff, Vincent M. Christoffels, Wil T. Labruyère, Antoon F. M. Moorman, Wouter H. Lamers

       Pages 185-197

   13. Effect of Cis-Located Human Satellite DNA on Electroporation Efficiency

       • Djenann Saint-Dic, Michael S. DuBow

       Pages 199-210

   14. Quantitation of Transient Gene Expression

       • Michael K. Showe, Louise C. Showe

       Pages 211-224

   15. Stable Integration of Vectors at High Copy Number for High-Level Expression in Animal Cells

       • James Barsoum

       Pages 225-237

   16. Electroporation of Drosophila Embryos

       • K. Puloma Kamdar, Thao N. Wagner, Victoria Finnerty

       Pages 239-243

The ability to introduce macromolecules into animal cells, includ­ ing DNA, RNA, proteins, and other bioactive compounds has facili­ tated a broad range of biological studies, from biochemistry and biophysics to molecular biology, cell biology, and whole animal stud­ ies. Gene transfer technology in particular will continue to play an essential role in studies aimed at improving our understanding of the relationships between the gene structure and function, and it has impor­ tant practical applications in both biotechnology and biomedicine, as evidenced by the current intense interest in gene therapy. Although DNA and other macromolecules may be introduced into cells by a variety of methods, including chemical treatments and microinjection, el- troporation has proven to be simpler to perform, more efficient, and effective with a wider variety of cell types than other techniques. The early and broad success of electric field-mediated DNA transfer soon prompted researchers to investigate electroporation for transferring other types of molecules into cells, including RNA, enzymes, antibodies, and analytic dyes. Animal Cell Electroporation and Electrofusion Protocols begins with three chapters that describe the theoretical and practical aspects of electroporation, including a review of the commercially available instrumentation. These introductory chapters will be of particular inter­ est to those new to electric field technologies and to those developing protocols for as yet untested species or cell types. Nineteen chapters follow that present well-tested protocols for electroporation of proteins and DNA into insect, fish, and mammalian cells.

"This volume should fulfill its aim, to "take the guesswork out of experimental trials." -Society for General Microbiology Quarterly

"The book is very up-to-date, for instance it cover the most important recent developments in experiments with isolation of tumor suppressor genes in melanoma by cDNA microarray. The book represents an outstanding work, which should not be missed in all laboratories dealing with melanoma as well as in all departments where melanoma patients are diagnosed and treated. Those people who are interested to learn how the completion of the Human Genome Project significantly has been contributing to new progress in cancer management, exampled by human malignant melanoma, will certainly enjoy reading this publication as well." - Neoplasma

• Department of Cancer Biology, Harvard University School of Public Health, Boston

  Jac A. Nickoloff

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