Optimum Conditions for Electric Pulse-Mediated Gene Transfer to Mammalian Cells in Suspension

A pulse-generating machine which delivers exponentially decaying pulses over broad range of pulse lengths was used to determine the optimum pulse conditions for gene transfer to FM3A cells. In the transformation of tk- cells with pTK1, a single pulse of 100-2000 μs gave a high transformation frequen...

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Veröffentlicht in:	Cell Structure and Function 1987, Vol.12(2), pp.173-180
Hauptverfasser:	Hama-Inaba, Hiroko, Takahashi, Mitsuko, Kasai, Michiki, Shiomi, Tadahiro, Ito, Akemi, Hanaoka, Fumio, Sato, Koki
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Biological and medical sciences Biotechnology Cell Line Cell Survival Cloning, Molecular Electric Stimulation Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Methods. Procedures. Technologies Mice Plasmids Thymidine Kinase - deficiency Thymidine Kinase - genetics Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Zusammenfassung:	A pulse-generating machine which delivers exponentially decaying pulses over broad range of pulse lengths was used to determine the optimum pulse conditions for gene transfer to FM3A cells. In the transformation of tk- cells with pTK1, a single pulse of 100-2000 μs gave a high transformation frequency at 1.5-6 kV/cm and room temperature, the highest transformation frequency obtained being 3 x 10-3. As the suspension buffer for cells exposed to the pulse, Saline G was better than PBS(-)for obtaining a large number of transformants because it ensured high cell viability.
ISSN:	0386-7196 1347-3700
DOI:	10.1247/csf.12.173