Tonya J. Webb Author Answered Apr 9, 2013
Microbiology and Immunology Department, University of Maryland School of Medicine, USA
Dear Antonia,
Please see below for a point by point response to your questions:
1. Yes, we collect blood from healthy donors.
2. No, the donors do not need to received G-CSF.
3. Without leukapheresis, the minimum amount of blood that I have used is from 2 CPT Vacutainer tubes (16-18ml).
4. If the initial isolation is skipped then the expansion will be minimal unless the donor has a high percentage of circulating NKT cells (>1% of the total T cell population).
5. I can usually keep culturing and expanding for NKT cells 5-6 weeks, but some lines have been maintained for ~3 months without thawing a fresh aliquot.
6. Yes, there is a high amount of variability between the donors, usually the higher the initial population of NKT cells the higher the expansion efficiency.
Good luck with your studies and please let me know if you have any additional questions.
Kind regards,
Tonya Webb