Preclinical ex vivo expansion of peripheral blood CD34+ selected cells from cancer patients mobilized with combination chemotherapy and granulocyte colony-stimulating factor

Authors: Lorenzon, D.¹; Mazzucato, M.²; Abbruzzese, L.²; Cilli, M.³; De Angeli, S.⁴; Degan, M.¹; Mambrini, G.⁵; Piccardi, F.³; Rupolo, M.⁶; Michieli, M.⁶; De Marco, L.²; Gattei, V.¹; Astori, G.

Source: Vox Sanguinis, Volume 94, Number 4, May 2008 , pp. 342-350(9)

Publisher: Wiley-Blackwell

Abstract:

Background and Objectives 

Ex vivo peripheral blood progenitor cell (PBPC) expansion has been proposed as a strategy to increase the number of haematopoietic progenitors available for cell transplantation. We have expanded CD34+ cells from PBPCs obtained from four patients with haematological malignancies and one patient with an Ewing's sarcoma. Materials and Methods 

Cells were expanded in the Dideco `Pluricell system'. After 12 days in culture, we evaluated cell phenotype, total nucleated cells, CD34+ fold increase, cell apoptosis and colony assay of expanded cells. Cell engraftment has been evaluated by transplanting two groups of irradiated non-obese diabetic/severe combined immunodeficient (NOD-SCID) mice with expanded and non-expanded cell populations. Results 

Total nucleated cells and CD34+ cells increased 59·5 and 4·0 times, respectively. The expanded cells were mainly constituted of myeloid and megakaryocytic cells. A significant increase in the number of colony-forming unit-granulocyte macrophage (CFU-GM) was observed in the CFU assay. Ten mice transplanted with expanded cells showed a best overall survival (80%) compared to 10 mice transplanted with non-expanded cells (20%). Human CD45+ cells were detected by flow cytometry and polymerase chain reaction in bone marrow and spleen of transplanted animals. The relative low engraftment level obtained with the expanded cells suggests a loss of SCID repopulating cells maybe due to cell differentiation during expansion. Conclusions 

We have demonstrated the feasibility of the ex vivo expansion of mobilized PBPCs from cancer patients, evidencing a clonal expansion of CFUs and the ability of the expanded cells to engraft the bone marrow and spleen of immunosuppressed mice. The differentiation of the CD34+ stem cell compartment could be further minimized by ameliorating the expansion conditions.

Keywords: ex vivo expansion bioreactors; haematopoietic stem cell transplantation; peripheral blood

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1423-0410.2008.01038.x

Affiliations: 1: Clinical and Experimental Haematology Research Unit, 2: Blood Bank and Department of Clinical Pathology and Immunohaematology, and 3: Animal Facility, Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy 4: Cell Culture Laboratory, Blood Bank, Treviso Regional Hospital, Treviso, Italy 5: Sorin Group Italia Srl Mirandola, MO, Italy 6: Medical Oncology, Centro di Riferimento Oncologico, IRCCS, Aviano, PN, Italy

Publication date: 2008-05-01

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• In this Subject: Therapeutics & Alternative Medicine
• By this author: Lorenzon, D. ;  Mazzucato, M. ;  Abbruzzese, L. ;  Cilli, M. ;  De Angeli, S. ;  Degan, M. ;  Mambrini, G. ;  Piccardi, F. ;  Rupolo, M. ;  Michieli, M. ;  De Marco, L. ;  Gattei, V. ;  Astori, G.

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