I have been using the Lonza nucleofector for RNP delivery to OT-I T cells without any issues for the past six months. However, recently, my nucleofected OT-I T cells have not been proliferating in vivo after LmOVA infection. It seems that nucleofected OT-I T cells can still proliferate in response to in vitro CD3/CD28 stimulation. Here are the conditions I am using:
- Cell type: Naive OT-I T cells
- Buffer: Lonza P4 nucleofection buffer
- Cell number: 1-5 x 10^6
- Nucleofection program: DS-189
What is the possible cause of this and how can I solve this problem?
Thank you in advance!
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