procedure is as follows: 
Collection of primate bone marrow cells 
Bone marrow cells are collected aseptically from the shaved 
betadine-treated iliac and/or long bones of primates anaesthetized with 
ketamine. Cells are aspirated using a bone marrow needle into syringes 
containing 1 ml of preservative-free porcine Intestinal heparin. Pooled 
samples of approximately 15 ml are layered over 25 ml of lymphocyte separation 
medium (LSM R ), or other marrow cell separation medium. In a 50 ml conical 50 ml 
polypropylene tube and centrifuged at 2000 x g for 15 minutes at 20°C. Cells 
at the Interface are collected, mixed with several volumes of Dulbecco's 
phosphate buffered saline (PBS), counted, pelleted (800 x g for 15 minutes at 
20°C). and resuspended to a volume corresponding to approximately 5 x 10 7 
nucleated cells/ml. 
Incubation of primate bone marrow cells with retroviral vector supernatant 
RVS preparations are thawed rapidly in a 37®C waterbath. Prepared bone 
marrow cells (BMC) are added to the RVS at a ratio of 5-10 vector particles/BMC 
and a polycation, either 8 ug/ml (final concentration) polybrene In early 
procedures or 5 ug/ml protamine in subsequent testing, is added. Tubes are 
tightly capped and returned to the water bath. After two hours incubation, the 
cells are pelleted at 800 x g for 10 minutes at 20°C, washed twice in PBS. and 
then resuspended in 10 ml PBS plus 202 autologous serum at a density of 3-6 x 
10 7 cell s/ml. The autologous serum is prepared from a sample of blood taken 
from the monkey at the time of marrow collection. The suspended cells are 
Infused back into the irradiated donor animal at 1 ml /minute. 
(2) Is the delivery systen efficient? What percentage of the 
target cells contain the added DNA? 
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Recombinant DNA Research, Volume 12 
