Murine allogeneic in vivo stem cell homing(,).
Stem cell homing has been studied in syngeneic models and appears to be rapid (<1 h) and dependent on cellular adhesion and migration factors. We utilized a full H2-mismatched transplantation model to determine the basics of allogeneic homing. C57BL/6J Lin-Sca-1+ cells were labeled with CFSE and injected in non-myeloablated BALB/c mice. Fluorescent cell detection was via high-speed FACS analysis. Alternatively, B6.SJL whole bone marrow cells were injected in lethally irradiated BALB/c mice (10 Gy). One, 3, 6, and 24 h after transplant, marrow was harvested and cells were either plated for high proliferative potential colony-forming cell (HPP-CFC) assay or secondarily injected into myeloablated (8 Gy) C57BL/6J mice using 10% competing C57BL/6J marrow. Chimerism was evaluated at 8 weeks. CFSE+ cells were detected in the bone marrow 1, 3, and 6 h after injection. The numbers were moderately lower when compared to syngeneic homing possibly due to strain effect. Conversely, utilizing a surrogate or secondary assay, we observed a decline of secondary engraftment of harvested cells over time, but not of HPP-CFC. Combining experiments and normalizing the 1-h time point to 100% (to allow comparison), we observed a mean relative engraftment of 87 +/- 29%, 72 +/- 21%, 84 +/- 35% of the 1 h level at 3, 6, and 24 h respectively. HPP-CFC assay showed no significant variation as a homing surrogate over 1-6 h. These data indicate a rapid homing into allogeneic recipients with a plateau at 1 h. The decline of secondary engraftability over time may indicate a phenotype alteration of homed cells.
Colvin
Gerald A
GA
Department of Research, Roger Williams Medical Center, Providence, Rhode Island, USA. gcolvin@lifespan.org
Lambert
Jean-Francois
JF
Dooner
Mark S
MS
Cerny
Jan
J
Quesenberry
Peter J
PJ
D000818:Animals; D000067696:Ataxin-1; D000067528:Ataxins; D001854:Bone Marrow Cells / Q000737:chemistry / Q000276:immunology / Q000502:physiology* / Q000528:radiation effects; D016026:Bone Marrow Transplantation; D019070:Cell Lineage; D002465:Cell Movement / Q000528:radiation effects; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006183:H-2 Antigens / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000276:immunology / Q000502:physiology* / Q000528:radiation effects; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D010641:Phenotype; D013388:Succinimides; D013997:Time Factors; D018183:Transplantation Chimera; D014184:Transplantation, Homologous; D014185:Transplantation, Isogeneic; D014916:Whole-Body Irradiation
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000067696:Ataxin-1; D000067528:Ataxins; C000593750:Atxn1 protein, mouse; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006183:H-2 Antigens; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D013388:Succinimides