Tissue Culture Shared ResourceEBV Sample Collection Information
A minimum of 6 ml of fresh blood in an ACD solution B tube ("yellow top tubes") is required.
The TCSR staff separate the peripheral white blood cells using Ficol and initiate transformation using the Epstein-Barr Virus. Backup blood samples are stored.
Cells are monitored closely until growth is observed. When growth is observed, cells are expanded.
When sufficient cells are available, 4 vials of 107 cells each are cryo-preserved. The vials are stored in redundant fashion as follows:
- 3 vials are stored in a LN2 vapor phase freezer.
- 1 vial is stored in a separate LN2 vapor phase freezer as a backup.
In the event that the transformation is not successful, the backup blood samples are used to attempt a new transformation. A total of 3 attempts will be made before declaring the transformation a failure.
As the stocks of transformed cells are depleted, the TCSR produces, tests, and stores new vials.
- Alternative tubes are not acceptable. Submit only ACD solution B tubes.
- Beware of "golden top tubes" which contain a clotting agent which renders samples useless.
- Samples must be received by the TCSR within 48 hours of draw. 24 hours is preferred.
- Samples should be held at room temperature until delivery.
- Do not refrigerate samples.
- Do not ship samples on wet ice.
- Samples must contain at least 6 ml of fresh blood.
If samples are received which do not meet the requirements, the researcher will be contacted.
The likelihood of success decreases significantly with failure to comply with the specifications.
If a transformation is attempted, the transformation fee will be charged, regardless of the success of the transformation.