An improved mouse Sca-1+ cell-based bone marrow transplantation model for use in gene- and cell-based therapeutic studies.
This study sought to develop a murine bone marrow transplantation strategy that would yield consistently high levels of long-term engraftment without significant morbidity and mortality. Hematopoietic stem cell (HSC)-enriched Sca-1+ cells were used for transplantation because of their propensity of homing to bone marrow. Green fluorescent protein (GFP)-expressing transgenic mice were used as donors. Murine Sca-1+ cells were enriched 13-fold from whole bone marrow with immunomagnetic column chromatography. Retroorbital injections yielded highly reproducible and higher levels of engraftment compared with tail vein injections. The combination of W41/W41 recipient mice and sublethal irradiation preconditioning produced long-term engraftment with minimal morbidity and mortality. A 24-hour delay between the sublethal irradiation and transplantation did not affect the efficiency and level of engraftment, but provided flexibility with respect to the timing of transplantation. Based on these findings, a mouse Sca-1+ cell-based strategy, involving the retroorbital injection of Sca-1+ cells into sublethally irradiated, myelosuppressed W41/W41 recipient mice within 24 h after irradiation, was developed. Transplantation of lentiviral vector-transduced wild-type Sca-1+ cells expressing GFP by this strategy led to consistently high levels of long-term engraftment. In summary, this murine Sca-1+ cell-based strategy could be used in studies of HSC-based gene or cell therapies.
Hall
Susan L
SL
Musculoskeletal Disease Center, Jerry L. Pettis Memorial VA Medical Center, Loma Linda, Calif. 92357, USA.
Lau
K-H William
KH
Chen
Shin-Tai
ST
Felt
Joshua C
JC
Gridley
Daila S
DS
Yee
Jiing-Kuan
JK
Baylink
David J
DJ
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis*; D016026:Bone Marrow Transplantation; D019070:Cell Lineage; D064987:Cell- and Tissue-Based Therapy; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology; D018189:Immunomagnetic Separation; D007275:Injections, Intravenous; D016086:Lentivirus / Q000235:genetics; D008565:Membrane Proteins / Q000032:analysis*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D023421:Models, Animal; D009915:Orbit / Q000098:blood supply; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D011828:Radiation Chimera; D011829:Radiation Dosage; D013623:Tail / Q000098:blood supply; D013997:Time Factors; D019172:Transplantation Conditioning / Q000379:methods; D014916:Whole-Body Irradiation / Q000379:methods
D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019009:Proto-Oncogene Proteins c-kit