title: Large volume ex vivo expansion of CD34-positive hematopoietic progenitor cells for transplantation.
abstract: A large volume culture system was developed for the ex vivo expansion of CD34 positive (+) hematopoietic progenitors, using cell 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-10 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-3/IL-6 significantly enhanced the expansion of myeloid progenitors, (c) the addition of IL-1 to SCF/IL-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.
authors.affiliation: University of Colorado Bone Marrow Transplant Program, USA.
authors.forename: M H
authors.initials: MH
authors.lastname: Purdy
authors.forename: C J
authors.initials: CJ
authors.lastname: Hogan
authors.forename: L
authors.initials: L
authors.lastname: Hami
authors.forename: I
authors.initials: I
authors.lastname: McNiece
authors.forename: W
authors.initials: W
authors.lastname: Franklin
authors.forename: R B
authors.initials: RB
authors.lastname: Jones
authors.forename: S I
authors.initials: SI
authors.lastname: Bearman
authors.forename: R J
authors.initials: RJ
authors.lastname: Berenson
authors.forename: P J
authors.initials: PJ
authors.lastname: Cagnoni
authors.forename: S
authors.initials: S
authors.lastname: Heimfeld
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans
chemical_list: D018952:Antigens, CD34