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Thrombopoietin to replace megakaryocyte‐derived growth factor: impact on stem and progenitor cells during ex vivo expansion of CD34+ cells mobilized in peripheral blood
Author(s) -
Duchez Pascale,
Chevaleyre Jean,
Vlaski Marija,
Dazey Bernard,
Bijou Fontanet,
Lafarge Xavier,
Milpied Noël,
Boiron JeanMichel,
Ivanovic Zoran
Publication year - 2011
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2010.02860.x
Subject(s) - megakaryocyte , thrombopoietin , progenitor cell , stem cell factor , cd34 , ex vivo , thrombopoiesis , stem cell , interleukin 3 , haematopoiesis , biology , chemistry , immunology , microbiology and biotechnology , in vivo , t cell , antigen presenting cell , immune system
BACKGROUND: The first protocol of ex vivo expansion that enabled almost total abrogation of postmyeloablative chemotherapy neutropenia was based on a three‐cytokine cocktail (stem cell factor [SCF], granulocyte–colony‐stimulating factor [G‐CSF], pegylated‐megakaryocyte growth and development factor [PEG‐MGDF]) in a serum‐free medium. Since the clinical‐grade molecule MGDF is no longer available on the market, we evaluated its substitution by thrombopoietin (TPO). STUDY DESIGN AND METHODS: CD34+ cells of myeloma patients were expanded for 10 days in serum‐free cultures with SCF, G‐CSF, or MGDF (100 ng/mL) or with TPO (2.5, 10, 20, 50, and 100 ng/mL) instead of MGDF. Day 10 amplifications of total nucleated cells, CD34+ cells, committed progenitors (CFCs), the capacity of engraftment of NOD/SCID mice (SCID repopulating cells [SRCs]), and the immunophenotype of cells in expansion product (CD13, CD14, CD33, CD41, CD61) were analyzed. RESULTS: TPO in doses of 2.5 and 10 ng/mL exhibits an effect comparable to that of MGDF (100 ng/mL) on total, CD34+, and CFCs amplification. Compared to MGDF, TPO (starting at 10 ng/mL) enhances two‐ to threefold the percentage of megakaryocyte lineage cells (CD41+ and CD61+). Finally, TPO maintains or even enhances (depending on dose) SRC activity. CONCLUSIONS: The use of TPO instead of MGDF in our protocol is feasible without any negative effect on progenitor cell expansion. Furthermore, applied in dose of 10 or 100 ng/mL it could enhance both the stem cell activity and the percentage of megakaryocyte lineage cells in expansion product.