Dendritic Cell Nucleofection
Dendritic Cell Nucleofection - Related Videos
11:39
T Cells Capture Bacteria by Transinfection from Dendritic Cells
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Cited by 2 •
2016
Here a protocol is presented to measure bacterial capture by CD4+ T cells which occurs during antigen presentation via transinfection from pre-infected dendritic cells (DC). We show how to perform the necessary steps: isolation of primary cells, infection of DC, DC/T cell conjugate formation, and measurement of bacterial T cell transfection.
10:24
Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
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Cited by 26 •
2011
Despite recent advancements in genetic modification, transfection of human embryonic stem cells (HESCs) remains a capricious process. To our knowledge, systematic and efficient methods to transfect human induced pluripotent stem cells (iPSCs) have not been reported. Here, we describe robust protocols to efficiently transfect and nucleofect human iPSCs.
09:04
Isolation of Mouse Lung Dendritic Cells
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Cited by 17 •
2011
A highly purified preparation of mouse lung dendritic cells is described. Specific emphasis is given to the isolation of conventional dendritic cell subset.
02:57
Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow
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2023
This video describes a protocol for the generation of dendritic cells--key antigen-presenting cells that activate T cells--from the bone marrow of an adult mouse.
09:07
Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus
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Cited by 27 •
2013
This protocol details a method to isolate antigen presenting cells from human thymus via different steps of enzymatic digestion of the tissue followed by density centrifugation of the single cell suspension and finally magnetic and/or FACS sorting of the cell populations of interest.
FAQs
The electrical pulse briefly changes membrane permeability, creating transient pores through which nucleic acids can enter. In the cell-type-specific solution, this pulse supports direct movement of DNA, RNA, or other nucleic acids beyond the membrane into the cytoplasm and nucleus. Because the permeability change is temporary, dendritic cells can subsequently recover and proceed to gene expression.
The cell-type-specific solution provides the chemical context in which the electrical pulse delivers nucleic acids to dendritic cells. Its use is linked to the specialized nature of the method, rather than treating all cell types identically. This compatibility supports the intended sequence of delivery, cellular recovery, and subsequent analysis of introduced nucleic acid activity.
Dendritic cell nucleofection can introduce DNA, RNA, or other nucleic acids, giving researchers more than one way to manipulate these immune cells. This flexibility supports controlled investigation of gene expression and cellular responses in dendritic cells. The resulting system can be adapted to questions involving antigen processing, cytokine responses, immune signaling, or interactions with infectious agents.
The central workflow consists of combining dendritic cells with the selected nucleic acid in the appropriate cell-type-specific solution, applying a brief electrical pulse, and allowing the cells to recover. The pulse promotes intracellular delivery, while the recovery period precedes gene expression. Researchers can then examine the resulting dendritic-cell response in the chosen experimental context.
This method is useful when researchers need controlled manipulation of primary or cultured dendritic cells. It can support experiments examining antigen processing, cytokine responses, host-pathogen interactions, and immune signaling. By introducing selected nucleic acids before analysis, investigators can connect altered cellular activity with specific immune processes rather than observing responses without experimental manipulation.
Manipulated dendritic cells can provide experimental systems for evaluating vaccine strategies and engineered cellular therapies, in addition to studying immune and infection biology. Their altered nucleic acid content enables analysis of how dendritic cells process antigens, produce cytokine responses, or participate in immune signaling. These outcomes help relate cellular manipulation to broader strategies for immune intervention.