Differential transduction efficiency of SCID-repopulating cells derived from umbilical cord blood and granulocyte colony-stimulating factor-mobilized peripheral blood.
The gene transfer efficiency into nonobese diabetic/severe combined immunodeficient (NOD/SCID)-repopulating cells (SRCs) derived from umbilical cord blood (UCB) (n = 11 NOD/SCID mice) and granulocyte-colony stimulating factor (G-CSF)-mobilized peripheral blood (MPB) (n = 64 NOD/SCID mice) was compared using a clinically relevant protocol and a retrovirus vector expressing the enhanced green fluorescent protein (EGFP). At 6-9 weeks after transplantation, the frequency of transduced human cells in the bone marrow (BM) (40.5% +/- 2.4% [mean +/- SE]) and spleen (SPL) (36.4% +/- 3.2%) in recipients of UCB cells was significantly higher (p < 0.001) than that observed in the BM (2.2% +/- 1.8%) and SPL (2.0% +/- 2.6%) in recipients of MPB. In subsequent studies, MPB was cultured for 2-8 days in cytokines prior to transduction to determine if longer prestimulation was required for optimal gene transfer. A significant increase in gene transfer into CD45(+) human cells and clonogenic cells derived from MPB SRCs was observed when cells were prestimulated for 6 days compared to 2 days prior to transduction (p = 0.019). However, even after 6 days of prestimulation, transduction was still significantly less than UCB. A substantial discrepancy exists in the ability to introduce genes effectively via retrovirus vectors into SRCs derived from MPB as compared to UCB.
Pollok
K E
KE
Herman B Wells Center for Pediatric Research, Riley Hospital for Children, Indianapolis, IN 46202, USA. kpollok@iupui.edu
van Der Loo
J C
JC
Cooper
R J
RJ
Hartwell
J R
JR
Miles
K R
KR
Breese
R
R
Williams
E P
EP
Montel
A
A
Seshadri
R
R
Hanenberg
H
H
Williams
D A
DA
D000818:Animals; D001773:Blood Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism* / Q000637:transplantation; D001803:Blood Transfusion; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016026:Bone Marrow Transplantation; D003114:Colony-Forming Units Assay; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D049452:Green Fluorescent Proteins; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000276:immunology; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics; D016511:Severe Combined Immunodeficiency / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics; D014181:Transplantation Immunology
D008164:Luminescent Proteins; D016179:Granulocyte Colony-Stimulating Factor; D049452:Green Fluorescent Proteins; D017493:Leukocyte Common Antigens