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EnglishA wide variety of mammalian cell types is used in gene transfection studies. Establishing transfection methods that enable highly efficient DNA uptake has become increasingly important. PC12 is an established rat pheochromocytoma cell line, which responds to exposure to NGF with cessation of growth, expression of cytoplasmic processes, and differentiation into cells resembling sympathetic neurons. Although PC12 cells represent an important model system to study a variety of neuronal functions, they proved relatively difficult to transfect. We have compared the efficiency of three different chemical transfection reagents (Lipofectamine 2000, Lipofectamine LTX and TransIT-LT1) and of two electroporation systems (Neon and Gene Pulser Xcell) in transiently transfecting undifferentiated PC12 cells. By comparing efficiencies from replicate experiments we proved electroporation (in particular Neon) to be the method of choice. By optimizing different parameters (voltage, pulse width and number of pulses) we reached high efficiency of transfection (90 %) and viability (99 %). We also demonstrated that, upon electroporation, cells are not altered by the transfection and maintain their ability to differentiate.
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| Titolo: | An electroporation protocol for efficient DNA transfection in PC12 cells |
| Autori: | Covello, Giuseppina; Siva, Kavitha; Adami, Valentina; Denti, Michela Alessandra |
| Autori Unitn: | |
| Titolo del periodico: | CYTOTECHNOLOGY |
| Anno di pubblicazione: | 2014 |
| Numero e parte del fascicolo: | 66 |
| Codice identificativo Scopus: | 2-s2.0-84904096728 |
| Codice identificativo Pubmed: | PMID: 23846478 |
| Codice identificativo ISI: | WOS:000339107800002 |
| Digital Object Identifier (DOI): | http://dx.doi.org/10.1007/s10616-013-9608-9 |
| Handle: | http://hdl.handle.net/11572/33156 |
| Appare nelle tipologie: | 03.1 Articolo su rivista (Journal article) |
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