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Selection and expansion of peripheral blood CD34+ cells in autologous stem
cell transplantation for breast cancer
SF Williams, WJ Lee, JG Bender, T Zimmerman, P Swinney, M Blake, J Carreon, M Schilling, S Smith, DE Williams, F Oldham and D Van Epps
Department of Medicine, University of Chicago Medical Center, IL, USA.
Cytopenia after high-dose chemotherapy and autologous stem cell reinfusion
is a major cause of morbidity. Ex vivo cultured expansion and
differentiation of CD34+ peripheral blood progenitor cells (PBPC) to
neutrophil precursors may shorten the neutropenic period further. We
explored the use of these ex vivo cultured PBPCs in nine patients with
metastatic breast cancer. All underwent PBPC mobilization with
cyclophosphamide, VP-16, and G-CSF. Subsequently, they underwent four to
five apheresis procedures. One apheresis product from each patient was
prepared using the Isolex 300 Magnetic Cell Separation System (Baxter
Immunotherapy, Irvine, CA) to obtain CD34+ cells. These cells were then
cultured in gas permeable bags containing serum-free X-VIVO 10
(BioWhittaker, Walkersville, MD) medium supplemented with 1% human serum
albumin and 100 ng/mL PIXY321. At day 12 of culture the mean fold expansion
was 26x with a range of 6 to 64x. One patient's cells did not expand
because of a technical difficulty. The final cell product contained an
average of 29.3% CD15+ neutrophil precursors with a range of 18.5% to
48.1%. The patients underwent high-dose chemotherapy with cyclophosphamide,
carboplatin, and thiotepa. On day 0, the cryopreserved PBPCs were reinfused
and on day +1 the 12-day cultured cells were washed, resuspended, and
reinfused into eight of nine patients. One patient was not infused with
cultured cells. The mean number of cultured cells reinfused was 44.6 x
10(6) cells/kg with a range of 0.8 to 156.6 x 10(6) cells/kg. No toxicity
was observed after reinfusion. The eight patients have recovered absolute
neutrophil counts > 500/microL on a median of 8 days (range 8 to 10
days); the median platelet transfusion independence occurred on day 10
(range 8 to 12 days) and platelet counts > 50,000/microL were achieved
by day 12 (range 9 to 14) for the seven patients whose platelet counts
could be determined. Expanded CD34+ selected PBPC can be obtained and
safely reinfused into patients.
Volume 87,
Issue 5,
pp. 1687-1691,
03/01/1996
Copyright © 1996 by The American Society of Hematology
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