5.0 STUDY DESIGN 
5.1 Patient Eligibility 
To ensure sufficient cells are present in the CD34 selected fraction as well as in 
the backup marrow at least 3 x 10* cells/kg must be harvested. If the yield is less 
than this number either a second harvest must be performed or the patient is not 
eligible for this study. 
5.2 Marrow Processing 
Details are provided in Appendix B. In brief mononuclear cells are obtained using 
the cell processor. 1/3 is frozen immediately without further manipulation and the 
remaining 2/3 is processed on the Cell Pro column to select CD34 cells as 
described in Appendix B. 
5.3 Transduction with the neomycin resistance gene 
CD34 selected cells are transduced with the GlNa or LNL6 vector, used in the 
previous St. Jude gene marking protocols, AMLREM, NEBREL, NEBREM, 
PAMMYLA and ETNA as described in Appendix C. The efficiency of marking 
will be estimated by clonogenic growth in G418 and semi-quantitative PCR for 
the neomycin resistance gene. 
5.4 Incubation with growth factors 
After marking one aliquot will be frozen immediately while the other aliquot will 
be incubated with the growth factors IL6, IL3 and SCF for 5-7 days as described 
in Appendix D. 
6.0 PATIENT ELIGIBILITY (See Section 1.0) 
6.1 All patients receiving an unpurged autologous BMT will be eligible for this 
protocol including :- 
6.1.1 Patients enrolled on the ABMT4 study or receiving unpurged marrow on 
the NB91 study (Dr Santana Principal Investigator) 
6.1.2 Patients receiving autografts on CNS 15 as part of therapy for brain 
tumors (Dr Heideman Principle Investigator) 
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Recombinant DNA Research, Volume 19 
