Blood in yellow top ACD tubes MUST be stored at room temperature until delivery. If at least 5 mL of blood is received, half of the sample will be used for the lymphocyte transformation and the other half will be used for isolation of one (1) 1 mL aliquot of frozen lymphocytes. If less than 5 mL of blood is received, the entire sample will be used for the lymphocyte transformation experiment.
The success rate for lymphocyte transformation depends on a number of factors. One key factor is the length of time between the blood draw and the transformation experiment.
We achieve a >95 percent success rate if the transformation is set up within 24 hours of phlebotomy. The success rate falls if more time elapses and is not usually successful if it is set up > 5 days from the time of phlebotomy. If the first transformation is not successful, we will set up a second EBV transformation with the second frozen lymphocyte sample that was reserved for this purpose. Please notify us if you do not want the second sample used as a "back-up" sample for transformation. Success rates for lymphocyte transformation range from 0–75 percent for samples that were previously isolated and frozen.
Lymphocyte transformation success also depends upon the number and condition of the starting cells, the EBV virus titer, and culturing techniques. We CANNOT ensure that lymphocyte transformation will be successful for every sample submitted to the HGCL core.
Transformed lymphocytes will be returned to the investigator in one of three ways:
- Three (3) frozen 1ml aliquots stored at -80oC (short term) and then transferred to -196oC in a liquid nitrogen freezer for long-term storage
- T25 flask of cells (~10ml culture) plus three (3) frozen 1ml aliquots
- T75 flask of cells (~40ml culture) plus three (3) frozen 1ml aliquots