Transgenic marrow transplantation: a new in vivo and in vitro system for experimental hemopoiesis and radiobiology which employs sequential molecular monitoring of multiple genetic markers.
Efforts to understand the in vivo regulation of hemopoiesis have been greatly impeded by an inability to trace and quantify the fate of multiple hemopoietic stem cell (HSC) cohorts within a single animal simultaneously. We report here the development of a molecular marker system which overcomes this deficiency and can be readily extended to a wide variety of applications in experimental hematology and radiobiology. Mixtures of HSC from multiple strains of transgenic mice, derived on a common genetic background, were used as donors to reconstitute lethally irradiated wild-type mice. Using molecular hybridization and the polymerase chain reaction, we documented the presence and quantified the amount of various transgenic markers in the blood of individual recipient mice after transplantation. The transgenic markers were also detected in spleen, thymus, peritoneal exudates, bone marrow, spleen colonies (CFU-S), and in vitro colonies. The transgenic transplantation marker system thus permits repeated analysis of multiple stem cell cohorts over the entire spectrum of hemopoiesis including HSC, intermediate precursors, and functionally mature cells. Therefore, the transgenic markers should facilitate the in vivo analysis of stem cell development, gene transfer, leukemogenesis, recovery from radiation or drug treatment, and the influence of hormones/growth factors on these processes.
Sigounas
G
G
Department of Experimental Hematology, Armed Forces Radiobiology Research Institute, Bethesda, Maryland 20892.
MacVittie
T J
TJ
D000818:Animals; D001483:Base Sequence; D016026:Bone Marrow Transplantation; D004247:DNA; D005240:Feasibility Studies; D005819:Genetic Markers; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000528:radiation effects; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D005819:Genetic Markers; D004247:DNA