CD19 CAR mRNA (Human)

Guide (The following is our recommend protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Primary T-cell culturing
1.1 Separate primary human T cells from fresh venous primary blood using LympPrep gradient centrifugation.
1.2 Purify to 95% purity.
1.3 Keep in liquid nitrogen prior to use.
1.4 Thaw and culture cells in complete culture medium for 24 h.
1.5 Activate with Human T-Activator CD3/CD28 Dynabeads (1:1 cell to bead ratio), and incubate for 48 h.
1.6 Remove Human T-Activator beads by gently pipetting up and down then transfer to a magnetic tube rack for 1-2 min, transfer supernatant containing cells to a fresh tube.
1.7 Electroporate the cells within 24 h of bead removal.
2. mRNA transfection
2.1 Add 50 ng/μL for each system unless specified otherwise (50 ng per reaction for triDrop, 500 ng per reaction for Neon, and 1000 ng per reaction for Lonza).
2.2 tore CD19 CAR mRNA at -80 °C in aliquots
3. Electroporation
3.1 Electroporate with triDrop Electroporation, Neon Electroporation or Lonza Electroporation protocol.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.