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Blood for the Establishment of Human B-Lymphoblastic Cell Lines Using Epstein-Barr Virus

Jeanne M. Meck, Ph.D., Director of Cytogenetics
Cytogenetics Laboratory
Institute for Molecular and Human Genetics
Georgetown University Hospital, 4th floor main, M4000
202-687-8810/202-687-3545 (weekdays)

Bassem Haddad, MD, Director of Tissue Culture Shared Resource
Victoria A. North, Manager of Tissue Culture Shared Resource
Lombardi Comprehensive Cancer Center
The Research Building
3rd floor, Room W314
202-687-0240 (weekdays)

1. Peripheral blood for EBV transformation is accepted Monday through Friday 8:30 a.m. - 4:30 p.m. except for certain holidays. Please provide as much notice as possible that samples will be coming so that we can ensure the minimum delay in starting to process the blood. Ideally, blood should be drawn in the morning so that it can be delivered to the cytogenetics lab by 1 p.m.. Samples arriving late in the day may not be processed until the following day which may adversely impact the probability of generating a viable cell-line from the blood.
2. The specimen should be drawn on the same day that it is delivered to the Cytogenetics Laboratory. However, blood drawn late in the day will usually be acceptable if transported to the lab the next morning. The chance of successfully generating a viable cell line from blood that is greater than 24 hours old is reduced.
3. It is essential that sterile technique be used when obtaining the blood.
4. Collect 5-10 ml of blood in a yellow top (Citrate) vacutainer tube. If possible use a 21 G needle. The use of smaller needles can increase the chance of cell lysis. Do not pop the top of the tube. Invert the tube six times immediately after it is drawn to ensure that the blood is well mixed with the anticoagulant to prevent clotting. We will attempt to isolated and culture cells from blood samples less than 5 ml, however, the chance of success decreases with the volume of blood we get.
5. Send blood to the cytogenetics lab immediately. Blood should be kept at room temperature. Do not refrigerate the blood even if it will not be delivered to the lab until the next day. Protect from heat and cold during shipping.
6. Please make sure that an EBV transformation request form accompanies the blood sample. This form should list the name of the referring physician, the name and date of birth of the patient and the time and date that the blood was drawn. Any additional clinical history can also accompany the sample.