Nanoelectroporation and Collection of Genetically Modified Exosomes in Primary Cultures of Dendritic Cells

Methods Mol Biol. 2020:2050:79-84. doi: 10.1007/978-1-4939-9740-4_8.

Abstract

Dendritic cells (DCs) are cells of the immune system that behave as antigen presenters and assist in T cell activation. DCs have recently been used in cell-based immunotherapies for the treatment of different diseases due to the lack of adverse nonspecific immune responses, typically elicited by other approaches. Genetically modified DCs, for example, have been used to stimulate CD4/CD8 antigen presenting immune responses. However, genetic manipulation of primary DCs remains a challenge. Here we describe a protocol for nonviral, benign transfection of primary DCs using nanochannel-based electroporation, and the subsequent collection of genetically modified exosomes for downstream applications.

Keywords: Cell-based immunotherapies; Dendritic cells; Nanochannel-based electroporation.

MeSH terms

  • Animals
  • Antigen Presentation
  • CD4 Antigens / metabolism
  • CD8 Antigens / metabolism
  • Cell Survival
  • Cells, Cultured
  • Dendritic Cells / cytology*
  • Dendritic Cells / immunology
  • Electroporation / instrumentation*
  • Exosomes / genetics*
  • Exosomes / transplantation
  • Humans
  • Mice
  • Nanotechnology
  • Primary Cell Culture / methods*
  • Transfection / instrumentation*

Substances

  • CD4 Antigens
  • CD8 Antigens