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t-cell-guide
Human Primary T cells: A Practical Guide
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Human Primary T Cells: A Practical Guide
Guide
Live document:
Human Primary T Cells: A Practical Guide
Preprint (v1):
https://peerj.com/preprints/26993/
Protocols:
https://www.protocols.io/view/human-primary-t-cells-a-practical-guide-quvdww6
Notebooks
Hundreds of millions of T cells can be isolated from a buffy coat sample
The relative abundance of T subsets vary across donors
Activation is required to rapidly expand T cells in culture but cell counts barely increase throughout the activation process
Electroporation is a feasible method for genetically manipulating T cells
Plasmid electroporation is inefficient and causes major cell death
mRNA electroporation is convenient for transiently expressing exogenous proteins
Cas9 RNP electroporation is a feasible technique to knockout a gene
Common T cell techniques can be optimized for cost
Untouched T cell isolation kit contents can be titrated down without major efficiency loss
Using fewer anti-CD3 and -CD28 beads for activation doesn’t immediately compromise activated cell yield
IL2 supplement is not necessary for short-term T cell cultures during and after CD3/CD28-based activation
T cell isolation can be circumvented by culturing PBMCs with activation beads
NIH’s reagent service provides access to highly standardized and cost-effective recombinant IL2