Direct comparison of steady-state marrow, primed marrow, and mobilized peripheral blood for transduction of hematopoietic stem cells in dogs.
The optimal stem cell source for stem cell gene therapy has not been defined. Most gene transfer studies have used peripheral blood or marrow repopulating cells collected after administration of granulocyte colony-stimulating factor and stem cell factor (G-CSF/SCF). For clinical applications, however, growth factor administration may not be feasible. Thus, in the current study we used a competitive repopulation assay in the dog to directly compare transduction efficiency of steady-state marrow, G-CSF/SCF-primed marrow, and G-CSF/SCF-mobilized peripheral blood. Cells from all three sources were transduced, cryopreserved, and thawed together before infusion into myeloablated dogs. Gene marking in hematopoietic repopulating cells was assessed by polymerase chain reaction. While primed marrow resulted in the highest long-term marking levels, steady-state marrow was transduced at least as efficiently as mobilized peripheral blood in all three dogs. These results suggest that steady-state marrow may be an appropriate source for genetic modification of hematopoietic stem cells.
Thomasson
Bobbie
B
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
Peterson
Laura
L
Thompson
Jesse
J
Goerner
Martin
M
Kiem
Hans-Peter
HP
D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D001853:Bone Marrow / Q000473:pathology*; D001854:Bone Marrow Cells / Q000166:cytology; D004285:Dogs; D018014:Gene Transfer Techniques; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D006133:Growth Substances / Q000378:metabolism; D006410:Hematopoiesis; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics; D019089:Stem Cell Factor / Q000378:metabolism; D033581:Stem Cell Transplantation; D013997:Time Factors
D018952:Antigens, CD34; D006133:Growth Substances; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor