Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system

Citation
H. Kawada et al., Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system, EXP HEMATOL, 27(5), 1999, pp. 904-915
Citations number
31
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Cardiovascular & Hematology Research
Journal title
EXPERIMENTAL HEMATOLOGY
ISSN journal
0301-472X → ACNP
Volume
27
Issue
5
Year of publication
1999
Pages
904 - 915
Database
ISI
SICI code
0301-472X(199905)27:5<904:REVEOH>2.0.ZU;2-1
Abstract
Cell numbers limit the widespread clinical use of cord blood (CB) for gene therapy and marrow replacement in adults; a simple and effective method for ex vivo expansion of CB primitive progenitor cells (PPC) is required, Rece ntly, the combination of thrombopoietin (TPO) and Flk-2/Flt-3 ligand (FL-2) was reported to support slow proliferation of CB-PPC in stroma-free liquid culture. We established a novel culture system in which the murine stromal cell line HESS-5 dramatically supports the rapid expansion of cryopreserve d CB-PPC in synergy with TPO/FL-2, Furthermore, while HESS-5 cells directly adhered to human progenitors during culture, the cultured human cells coul d easily be harvested without contamination by HESS-5 cells. Within 7 days of culture, a 100-fold increase in CD34(bright)/CD38(dim) cells was obtaine d in serum-containing culture, When HESS-5 cells were physically separated from human progenitor cells in the presence of TPO/FL-2, synergy was blocke d, suggesting that HESS-5 cells support proliferation of PPC by direct cell -to-cell interaction, The hematopoietic-supportive effects of this xenogene ic coculture system were then assessed in a very short-term (5 days) serum- free culture, Expansion was further enhanced by addition of stem cell facto r (SCF) or interleukin-3 (IL-3), As a result, a 50- to 100-fold increase in CD34(bright)/CD38(dim) cells was noted, Colony-forming units in culture (C FU-C) and mixed colonies (CFU-GER IRI) were enhanced by 10- to 30-fold and 10- to 20-fold, respectively, Moreover, generation of long-term-culture-ini tiating cells (LTC-IC) from CD34(bright)/CD38(dim) cells was amplified by 2 5-fold, The severe-combined immunodeficient (SCID) mouse-repopulating cell (SRC) assay confirmed extensive ability of the expanded cells to reconstitu te long-term hematopoiesis. These results indicate that this xenogeneic coc ulture system, in combination with human cytokines, can rapidly generate PP C from cryopreserved CB, (C) 1999 International Society for Experimental He matology. Published by Elsevier Science Inc.