Recombinant Sendai virus provides a highly efficient gene transfer into human cord blood-derived hematopoietic stem cells.
Hematopoietic stem cells (HSCs) are a promising target for gene therapy, however, the low efficiencies of gene transfer using currently available vectors face practical limitations. We have recently developed a novel and efficient gene transfer agent, namely recombinant Sendai virus (SeV), and we have here characterized SeV-mediated gene transfer to human cord blood (CB) HSCs and primitive progenitor cells (PPC) using the jelly fish green fluorescent protein (GFP) gene. Even at a relatively low titer (10 multiplicity of infections), SeV achieved highly efficient GFP expression in CB CD34(+) cells (85.5+/-5.8%), as well as more immature CB progenitor cells, CD34(+)AC133(+) (88.2+/-3.7%) and CD34(+)CD38(-) (84.6+/-5.7%) cells, without cytokines prestimulation, that was a clear contrast to the features of gene transfer using retroviruses. SeV-mediated gene transfer was not seriously affected by the cell cycle status. In vitro cell differentiation studies revealed that gene transfer occurred in progenitor cells of all lineages (GM-CFU, 73.0+/-11.1%; BFU-E, 24.7+/-4.0%; Mix-CFU, 59+/-4.0%; and total, 50.0+/-7.0%). These findings show that SeV could prove to be a promising vector for efficient gene transfer to CB HSCs, while preserving their ability to reconstitute the entire hematopoietic series.
Jin
C H
CH
Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Kusuhara
K
K
Yonemitsu
Y
Y
Nomura
A
A
Okano
S
S
Takeshita
H
H
Hasegawa
M
M
Sueishi
K
K
Hara
T
T
D000818:Animals; D003114:Colony-Forming Units Assay; D036101:Cord Blood Stem Cell Transplantation; D005312:Fetal Blood / Q000166:cytology*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D029082:Sendai virus / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins