Author:

Keywords:

Animals, Antigens, CD34, Cell Division, Cell Line, Cell Lineage, Coculture Techniques, Cytokines, Fetal Blood, Hematopoiesis, Hematopoietic Stem Cell Mobilization, Humans, Killer Cells, Natural, Mice, Stem Cells, Time Factors, Science & Technology, Life Sciences & Biomedicine, Hematology, Medicine, Research & Experimental, Research & Experimental Medicine, hematopoiesis, AFT024, expansion, multi-lineage, cord blood, LONG-TERM CULTURE, COLONY-STIMULATING FACTOR, EX-VIVO EXPANSION, LIMITING DILUTION ASSAYS, MURINE BONE-MARROW, KILLER NK CELLS, STEM-CELLS, IN-VITRO, INITIATING CELLS, SELF-RENEWAL, 1102 Cardiorespiratory Medicine and Haematology, Immunology, 3201 Cardiovascular medicine and haematology

Abstract:

OBJECTIVE: The majority of studies assessing ex-vivo expansion of primitive hematopoietic cells only address production of myeloid progeny whereas it may be more appropriate to maintain or expand progenitors that retain capacity for multilineage differentiation. In this study, we assessed the capacity of the murine fetal liver cell line AFT024 to expand primitive myeloid progenitors (LTC-IC) and lymphoid progenitors (NK-IC) from umbilical cord blood (CB) and mobilized peripheral blood (PB) CD34(+)lin(-)38(-) cells. METHODS: Sorted cells were established in expansion cultures in direct contact with the feeder or in a transwell above the feeder (noncontact culture) and various combinations of Flt-3L (FL), stem cell factor, interleukin 7, thrombopoietin (Tpo), and macrophage inflammatory protein-1alpha added. Frequency of LTC-IC and NK-IC was assessed at day 0 and following 2 and 5 weeks expansion culture. RESULTS: CB contained significantly more LTC-IC and NK-IC at day 0 and showed an enhanced capacity for expansion compared to PB. The combination of FL and Tpo showed maximal expansion of CB LTC-IC and NK-IC at 5 weeks in both contact and noncontact conditions. In contrast, expansion of PB LTC-IC and NK-IC was maximal at 2 weeks and required multiple cytokines. CONCLUSIONS: These results demonstrate that AFT024 can expand primitive hematopoietic progenitors from CB and PB and expanded cells retain the capacity for myeloid and lymphoid differentiation. These findings emphasize the importance of assessing multi-lineage differentiation capacity following ex-vivo expansion. Elucidation of specific factors necessary for ex-vivo expansion will contribute to the development of a clinically applicable system.