Mobilization of long-term hematopoietic reconstituting cells in mice by the combination of stem cell factor plus granulocyte colony-stimulating factor

Abstract

In this study, we have compared the ability of recombinant human granulocyte colony-stimulating factor (rhG-CSF) alone and the combination of low doses of recombinant rat pegylated stem cell factor (rrSCF-PEG) plus rhG-CSF to mobilize peripheral blood progenitor cells (PBPCs) with long-term engrafting potential. Female recipient irradiated mice were transplanted with PBPCs from male mice that were mobilized with rhG-CSF alone (group A) or rrSCF-PEG plus rhG-CSF (group B). As previously shown, greater short-term survival resulted in group B compared with group A, with 80% and 40% survival at 30 days posttransplant, respectively. Both groups of animals showed long-term donor-derived engraftment in greater than 95% of animals, as determined by quantitative specific polymerase chain reaction amplification of a Y chromosome sequence from whole blood of the mice at 6 to 12 months posttransplantation. Analysis of individual granulocyte-macrophage colonies, picked up from semisolid methylcellulose culture of bone marrow cells from transplanted mice, resulted in detection of donor- derived DNA in 98% of colonies from group B mice compared with 81% from group A mice. These data show that cells with long-term potential are mobilized by rhG-CSF alone and the combination of rrSCF-PEG plus rhG- CSF. Furthermore, an increased number of cells with short-term and long- term engraftment potential was obtained with rrSCF-PEG plus rhG-CSF compared with rhG-CSF alone.

Growth factor release from dentine matrix by pulp‐capping agents promotes pulp tissue repair‐associated events

“…, Yan et al . ) and is used therapeutically to enhance the release of haematopoietic stem cells as an adjunctive therapy in patients undergoing chemotherapy and/or radiotherapy. Chemotaxis of cells involved in tissue repair is a fundamental process in the healing of any tissue; with evidence emerging that that cells involved in pulp tissue regeneration may not, in fact, only originate from local tissue niches, but may be haematopoietic or perivascular in origin (Feng et al .…”

Mobilization of primitive haemopoietic progenitor cells and stem cells with long‐term repopulating ability into peripheral blood in mice by pegylated recombinant human megakaryocyte growth and development factor

“…A variety of growth factor combinations in PBPC mobilization have been tested in animal studies. For example, G‐CSF cooperates with SCF ( Briddell et al , 1993 ; Yan et al , 1994 ) or flt‐3 ligand ( Sudo et al , 1997 ; Molineux et al , 1997 ) to enhance PBPC mobilization. Our data show in vivo synergy between PEG‐rHuMGDF and rhG‐CSF in PBPC mobilization in mice.…”

“…To assess long‐term engraftment of the transplanted donor PB cells in female recipients, four mice that survived at approximately 6 months after transplantation were treated with 300 μg/kg/d of PEG‐rHuMGDF plus 500 μg/kg/d of rhG‐CSF for 5 d and then the mobilized PB was plated in the methylcellulose culture as described above. After 7 d of culture, up to 48 GM colonies derived from each mouse were individually picked from dishes and analysed by PCR for the presence of sequences specific for murine Y chromosome and murine PDGF B receptor as an internal standard, as previously reported ( Yan et al , 1994 ) with some modifications. Briefly, colonies were transferred into 150 μl of H 2 O in plastic tubes, boiled for 15 min, kept on ice for 5 min, and incubated with 2 μl of 14.4 mg/ml proteinase K (Boehringer Mannheim, Mannheim, Germany) at 60°C for 2 h. Following further boiling for 15 min, 15 μl of each resultant sample was used for PCR amplification.…”

“…Among several growth factors, granulocyte colony‐stimulating factor (G‐CSF) is an effective mobilizer of PBPCs ( Pojda et al , 1990 ; Molineux et al , 1990a , b; Dührsen et al , 1988 ) and therefore has already been applied to PBPC mobilization for clinical use. Other growth factors such as stem cell factor (SCF) ( Andrews et al , 1992 ; Bodine et al , 1993 ) and flt‐3 ligand ( Brasel et al , 1996 ) can also mobilize PBPCs and enhance PBPC numbers in synergy with G‐CSF ( Briddell et al , 1993 ; Yan et al , 1994 ; Sudo et al , 1997 ; Molineux et al , 1997 ).…”

Elevation of extracellular adenosine mobilizes haematopoietic progenitor cells and granulocytes into peripheral blood and enhances the mobilizing effects of granulocyte colony‐stimulating factor

“…Numerous experimental settings comprising combined administration of G‐CSF with various cytokines or growth factors, e.g. stem cell factor (4, 5), interleukin‐3 (6), FLT‐3 ligand (7, 8), or megakaryocyte growth and development factor (9) have been successfully tested. Nevertheless, in current clinical practice, G‐CSF is used most commonly as the only mobilization‐stimulating agent; only sometimes it is combined with cyclophosphamide (10).…”