cells are acquired (usually daily collections for 3-5 
days) . 
7 . 2 Marrow Harvest 
The technique for bone marrow harvesting has been 
described in detail. In brief, at the prescribed time, 
the patient is taken to the operating room where, under 
general anesthesia, multiple marrow aspirations from 
the anterior and posterior iliac crests are carried out 
under sterile conditions. Volumes totalling 1000-2500cc 
are collected into heparinized syringes and pooled in a 
heparinized Travenol bone marrow collection kit. 
Marrow is processed by passage through screens and 
samples are taken for counting and microbial cultures 
as well as specimens for clonogenic assays. The pooled 
marrow preparation is transferred to a 2 L 
Fenwal collection bag and taken immediately to the 
cryopreservation laboratory for processing, counts, 
cultures, and freezing. A minimum total number of 0.5 
x 10 8 marrow cells/kg is necessary to ensure proper 
engraftment. If less- than 0.5 x 10 8 nucleated cells/kg 
are obtained, then no high dose 
chemotherapy/TBI/autotransplantation will be attempted. 
If deemed appropriate by the principal investigator, a 
second marrow harvest may be attempted in order to 
obtain adequate numbers of cells. Maximum marrow 
procurement exceeding the minimum will be attempted in 
each case. 
7.3 Cryopreservation of Cells 
A solution containing 10% DMSO plus autologous plasma 
in McCoy's is added in equal amounts to the cell 
suspension that is kept on ice. Then the freezing bags 
containing the mixture are placed in a Cryo-Med Control 
Rate Freezer and cooled at a 1° C/minute down to -3 0° 
C. At that time, the freezing rate is increased to 
10° C. /minute until a temperature of -90° C is reached. 
Then the freezing bags are removed and placed into a 
liquid nitrogen storage freezer. 
8.0 Pre-study Evaluation (see Appendix H) 
8.1 Baseline evaluation prior to entry into the protocol 
Clinical Evaluation 
1. History and physical examination with height and 
weight and body surface calculations. 
2. Chest and other x-rays as clinically indicated. 
Recombinant DNA Research, Volume 16 
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