Improved hematopoietic stem cell engraftment following ex vivo expansion of murine marrow cells with SCF and Flt3L

Background In vitro incubation of murine BM cells with IL-3, IL-6, IL-11 and SCF induces expansion of HPC but fails to preserve ‘engraftability’ in comparison with normal untreated marrow cells. We studied how culturing marrow cells for 48 and 72 h with a combination of the cytokines SCF and Flt3L i...

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Published in:Cytotherapy (Oxford, England) England), 2007, Vol.9 (6), p.532-538
Main Authors: Diehl, A, Stoelting, S, Nadrowitz, R, Wagner, T, Peters, S.O., MD
Format: Article
Language:English
Subjects:
Advanced Basic Science
Animals
Bone Marrow Cells - cytology
Bone Marrow Cells - drug effects
Bone Marrow Transplantation
Cell Count
Cell Proliferation - drug effects
Cells, Cultured
Colony-Forming Units Assay
cytokines
engraftment
Hematopoietic Stem Cell Transplantation - methods
Interleukins - pharmacology
Male
Membrane Proteins - pharmacology
Mice
Mice, Inbred C57BL
Models, Biological
Other
stem cell expansion
Stem Cell Factor - pharmacology
Time Factors
Whole-Body Irradiation
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Summary:Background In vitro incubation of murine BM cells with IL-3, IL-6, IL-11 and SCF induces expansion of HPC but fails to preserve ‘engraftability’ in comparison with normal untreated marrow cells. We studied how culturing marrow cells for 48 and 72 h with a combination of the cytokines SCF and Flt3L influences cell expansion and engraftability. Methods Competitive repopulation of lethally irradiated C57BL/6 mice was used to examine engraftability of ex vivo cytokine-expanded Ptprc chimeric BM. A methylcellulose in vitro assay was used to determine the expansion of substitute progenitors. Results Both cytokine combinations successfully expanded progenitor populations when assayed in methylcellulose culture in vitro . After 72 h, the colony numbers of the expansion cultures increased 61% with IL-3, IL-6, IL-11 and SCF stimulation and 96% with SCF and Flt3L stimulation. Engraftment of competitively transplanted cells, cultured with IL-3, IL-6, IL-11 and SCF, consistently dropped to levels below 16%. However, 48 h culture with SCF and Flt3L resulted in 53.5 ± 1.6% engraftment at 17 days and 64 ± 3.7% engraftment at 19 weeks post-transplantation. Extending the cytokine exposure to 72 h resulted in 70 ± 4.4% short-term engraftment at 17 days, and 64 ± 2.4% engraftment at 19 weeks post-transplantation. Discussion The data demonstrate the ability of SCF and Flt3L cytokine-stimulated BM cells to maintain short- and long-term engraftability. We conclude that these cytokines play a crucial role in maintaining engraftment of hematopoietic progenitors.
ISSN:1465-3249
1477-2566
DOI:10.1080/14653240701452073