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Transfection and Transformation of Human Thyroid Epithelial Cells

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Basic Cell Culture Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 75))

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Abstract

High-efficiency gene transfer into mammalian fibroblasts is a routine procedure that can be performed by a variety of methods (1), but transfection of epithelial cells has been more difficult to achieve. Each of the standard techniques has associated problems; special equipment is required for electroporation (2) and also for direct microinjection (3). Protoplast fusion (4) is suitable only for cloned genes. Retroviral transduction of genes (5) is a method of high efficiency for animal cells, but again is applicable only to cloned genes of restricted size, and safety considerations are likely to restrict greatly the use of retroviral vectors suitable for human cells. Calcium phosphate coprecipitation has been used with success in several epithelial lines (68) and, more recently, strontium phosphate coprecipitation (9). These techniques are very simple to perform, require no special equipment, and can be applied to genomic DNA transfection.

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References

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© 1997 Humana Press Inc., Totowa, NJ

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Lemoine, N.R., Wynford-Thomas, D. (1997). Transfection and Transformation of Human Thyroid Epithelial Cells. In: Pollard, J.W., Walker, J.M. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-441-0:441

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  • DOI: https://doi.org/10.1385/0-89603-441-0:441

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-441-9

  • Online ISBN: 978-1-59259-561-7

  • eBook Packages: Springer Protocols

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