Expansion of LTC-ICs and Maintenance of p21 and BCL-2 Expression in Cord Blood CD34+/CD38 Early Progenitors Cultured over Human MSCs as a Feeder Layer

Authors

  • Suzanne Kadereit,

    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Linda S. Deeds,

    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Stephen E. Haynesworth,

    1. Department of Biology, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Omer N. Koc,

    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
    2. NCI Comprehensive Cancer Center, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Margaret M. Kozik,

    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Emese Szekely,

    1. NCI Comprehensive Cancer Center, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Kathleen Daum-Woods,

    1. NCI Comprehensive Cancer Center, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Glenn W. Goetchius,

    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Pingfu Fu,

    1. Department of Epidemiology and Biostatistics, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
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  • Lisbeth A. Welniak,

    1. SAIC Frederick, NCI at Frederick, Frederick, Maryland, USA
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  • William J. Murphy,

    1. SAIC Frederick, NCI at Frederick, Frederick, Maryland, USA
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  • Mary J. Laughlin M.D.

    Corresponding author
    1. Department of Medicine, Case Western Reserve University/University Hospitals of Cleveland, Cleveland, Ohio, USA
    • Director, Allogeneic Transplant Program, Case Western Reserve University, University Hospitals Ireland Comprehensive Cancer Center, 11100 Euclid Avenue, Cleveland, Ohio 44106-5061, USA. Telephone: 216-844-4766; Fax: 216-844-3616
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Abstract

Allogeneic transplantation with umbilical cord blood (UCB) is limited in adult recipients by a low CD34+ cell dose. Clinical trials incorporating cytokine-based UCB in vitro expansion have not demonstrated significant shortening of hematologic recovery despite substantial increases in CD34+ cell dose, suggesting loss of stem cell function. To sustain stem cell function during cytokine-based in vitro expansion, a feeder layer of human mesenchymal stem cells (MSCs) was incorporated in an attempt to mimic the stem cell niche in the marrow microenvironment. UCB expansion on MSCs resulted in a 7.7-fold increase in total LTC-IC output and a 3.8-fold increase of total early CD34+ progenitors (CD38/HLA-DR). Importantly, early CD34+/CD38/HLA-DR progenitors from cultures expanded on MSCs demonstrated higher cytoplasmic expression of the cell-cycle inhibitor, p21cip1/waf1, and the antiapoptotic protein, BCL-2, compared with UCB expanded in cytokines alone, suggesting improved maintenance of stem cell function in the presence of MSCs. Moreover, the presence of MSCs did not elicit UCB lymphocyte activation. Taken together, these results strongly suggest that the addition of MSCs as a feeder layer provides improved conditions for expansion of early UCB CD34+/CD38/HLA-DR hematopoietic progenitors and may serve to inhibit their differentiation and rates of apoptosis during short-term in vitro expansion.

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