required. 
B. Administration and Toxicides: 
(1) The dose of G-CSF will be 16 /xg/kg/day. The appropriate amount of G-CSF is 
drawn into 2-3 ml syringes containing approximately 1.5 - 2 ml each. G-CSF is 
injected subcutaneously in the thighs or abdomen at 2 sites using a 25-30 gauge 
needle. Toxicity will be judged using a standard format for toxicity grading as 
outlined in Appendix A. Toxicides previously encountered with G-CSF are mainly 
bone pain. If any donor develops Grade 2 or above toxicity the dose for the next 
donor will be lowered by 50%. Any donor who develops an absolute neutrophil 
count in excess of 50,000/mm 3 will have the dose of G-CSF halved on that day. 
G-CSF will be deleted if the absolute neutrophil count exceeds 75,000/mm 3 In on 
the next day the absolute neutrophil count is below 50,000/mm 3 the G-CSF will 
be resumed. 
C. PBSC Collections: 
(1) Subclavian Catheters: 
(a) Percutaneous subclavian catheter will be placed preferentially. Potential 
donors who refuse a central venous catheter will be evaluated by Dr. 
Bensinger, Buckner or Rowley for consideration of a vein to vein 
procedure. If the veins are adequate, apheresis will be carried out on a 
vein to vein basis. 
Potential complications related to the catheter include infection and 
clotting. Patients will be monitored daily for local inflammation and fever. 
Febrile episodes while not in the hospital require a prompt visit to the on- 
call physician. Episodes of clotting will be treated with urokinase as per 
the standard practice manual. 
(2) Continuous Flow Centrifugation: 
(a) PBSC’s are collected by continuous flow centrifugation using standard 
techniques established in our laboratory. This procedure separates cells 
continuously on the basis of differential sedimentation in a centrifuge (CS- 
3000 or Cobe 2997). Each collection takes 4 hours and involves 
heparinizing the donor (13,000 units over 4 hours), infusing approximately 
100 ml 6% hydroxyethyl starch and processing the donor’s blood at a high 
flow rate. To be practical on a daily basis, this flow rate requirement can 
best be met with the placement of a central venous double lumen catheter. 
Each days collection deprives the donor of approximately 300 ml of 
plasma, 50 ml of packed red cells, 30 X 10 9 leukocytes and 100 X 10 9 
platelets. The leukocytes, plasma, and platelets are rapidly replaced by the 
donor, but replacement of red cells is hampered by the removal of 
reticulocytes in the buffy coat and the donor’s hematocrit decreases about 
1 point daily. This should not pose a problem in donor’s who have not had 
marrow harvested. 
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Recombinant DNA Research, Volume 16 
