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Transfection of bone marrow derived cells with immunoregulatory proteins.
Khantakova, Julia N; Silkov, Alexander N; Tereshchenko, Valeriy P; Gavrilova, Elena V; Maksyutov, Rinat A; Sennikov, Sergey V.
Afiliación
  • Khantakova JN; Federal State Budgetary Scientific Institution "Research Institute of Fundamental and Clinical Immunology", Yadrintsevskaya St. 14, Novosibirsk 630099, Russia.
  • Silkov AN; Federal State Budgetary Scientific Institution "Research Institute of Fundamental and Clinical Immunology", Yadrintsevskaya St. 14, Novosibirsk 630099, Russia.
  • Tereshchenko VP; Federal State Budgetary Scientific Institution "Research Institute of Fundamental and Clinical Immunology", Yadrintsevskaya St. 14, Novosibirsk 630099, Russia.
  • Gavrilova EV; State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk 630559, Russia.
  • Maksyutov RA; State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk 630559, Russia.
  • Sennikov SV; Federal State Budgetary Scientific Institution "Research Institute of Fundamental and Clinical Immunology", Yadrintsevskaya St. 14, Novosibirsk 630099, Russia. Electronic address: sennikovsv@gmail.com.
Cytokine ; 108: 82-88, 2018 08.
Article en En | MEDLINE | ID: mdl-29579547
In vitro electroporation gene transfer was first performed in 1982. Today, this technology has become one of the major vehicles for non-viral transfection of cells. All non-viral transfections, such as calcium phosphate precipitation, lipofection, and magnetic transfection, have been shown to achieve a transfection efficiency of up to 70% in commonly used cell lines, but not in primary cells. Here we describe the use of electroporation to transfect primary mouse bone marrow-derived cells, such as macrophages (Mφ) and dendritic cells (DCs) with high efficiencies (45%-72%) and minimal cell death. The transfection efficiencies and cell death varied depending on the culture duration of the DCs and Mφ. Moreover, the electroporation efficiency was increased when conditioning medium was used for culturing the cells. Furthermore, we demonstrated that measuring the plasmid-encoded secreted proteins is a highly sensitive method for determining the transfection efficiency. In summary, electroporation with plasmid vectors is an efficient method for producing DCs and Mφ with transient expression of immunoregulatory proteins.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Células de la Médula Ósea / Transfección / Interleucina-10 / Electroporación / Receptores Tipo I de Factores de Necrosis Tumoral Límite: Animals Idioma: En Revista: Cytokine Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Rusia

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Células de la Médula Ósea / Transfección / Interleucina-10 / Electroporación / Receptores Tipo I de Factores de Necrosis Tumoral Límite: Animals Idioma: En Revista: Cytokine Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Rusia