Pre-IDE Submission: Clinical Protocol 
Neuroblastoma Bone Marrow Purging System 
BAXTER HEALTHCARE CORPORATION, HYLAND DIVISION 
Post-transplantation 
a) physical exam as per regime 
b) CBC with platelets and differential daily until self sustaining 
c) blood chemistries as per regime 
d) BM aspirate as clinically indicated for diagnosis and documentation of recovery, 
and at day 28-30, at day 60, and at 6 months for analysis of engraftment and 
presence of neogene 
6. 1.8. 2 Laboratory Assessment 
In addition to the above, in vitro studies which directly relate to evaluation of the 
purging/gene marker procedure will be performed as outlined below: 
In Vitro Marrow Testing 
The following tests will be performed on the marrow cell suspension immediately before 
and after transduction and immediately after the purging procedure: 
i) Total nucleated cell count 
ii) Immunofluorescent staining and cell sorting for detection and separation of 
neuroblasts 
iii) Immunocytologic analysis for the detection of neuroblasts 
iv) Clonogenic assay for neuroblasts and normal progenitor cells, analysis of 
clones for presence of the marker 
v) Sterility evaluation - aerobic and anaerobic liquid cultures & blood agar plates. 
vi) Nucleated cell viability by trypan blue exclusion 
vii) Packed cell volume/Hematocrit* 
* To be performed only before immunomagnetic purging 
Recombinant DNA Research, Volume 16 
