Large Volume Ex Vivo Expansion of CD34-Positive Hematopoietic Progenitor Cells for Transplantation
M.H. Purdy, C.J. Hogan, L. Hami, I. McNiece, W. Franklin, R.B. Jones, S.I. Bearman, R.J. Berenson, P.J. Cagnoni, S.Heimfeld, E.J. Shpall
JOURNAL OF HEMATOTHERAPY 4:515-525 (1995)
A large volume culture system was developed for the ex vivo expansion of CD34 positive (+) hematopoietic progenitors, using cells donated by 15 patients receiving high-dose chemotherapy with autologous hematopoietic progenitor cell support (AHPCS).
Substantial expansion of myeloid (181- fold) and megakaryocyte (41-fold) progenitors cells was demonstrated, using the conditions that we determined to be optimal: CD34+ progenitors cultured unperturbed for 7 (marrow) or 10 (blood) days in Teflon-coated bags with X-Vivo-l0 medium containing 10% autologous plasma, 100 ng/ml, respectively, of recombinant stem cell factor (SCF), interleukin 3 (IL-3), interleukin 6 (IL-6), and granulocyte colony-stimulating factor (G-CSF).
The studies demonstrated that
(a) CD34 selection was necessary to obtain large, clinically relevant numbers of hematopoietic progenitors,
(b) the addition of G-CSF to the baseline regimen of SCF/IL-31IL-6 significantly enhanced the expansion of myeloid progenitors,
(c) the addition of IL-l to SCFIIL-3/IL-6 did not significantly enhance myeloid progenitor cell expansion,
(d) CD34+ G-CSF-mobilized peripheral blood 'progenitor cells (PBPC) produced higher numbers of myeloid progenitors in culture than CD34+ marrow cells, and
(e) long-term tissue culture (LTC) assays demonstrate the preservation of long-term initiating cells in ex vivo culture.
The short-term and long-term reconstituting capability of CD34+ PBPC cultured in this system remains to be determined and will be evaluated in a clinical trial where they will be used as the sole source of AHPCS following high-dose therapy.
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