Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 2095-2108 |
| Number of pages | 14 |
| Journal | Human Gene Therapy |
| Volume | 12 |
| Issue number | 17 |
| DOIs | |
| State | Published - 2001 |
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ASJC Scopus subject areas
- Genetics
Cite this
Differential transduction efficiency of SCID-repopulating cells derived from umbilical cord blood and granulocyte colony-stimulating factor-mobilized peripheral blood. / Pollok, Karen; Hanenberg, Helmut; Williams, David A.; Van der Loo, Johannes C M; Cooper, Ryan J.; Hartwell, Jennifer R.; Miles, Kathryn R.; Breese, Robert; Williams, Emily P.; Montel, Angela; Seshadri, Roopa.
In: Human Gene Therapy, Vol. 12, No. 17, 2001, p. 2095-2108.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Differential transduction efficiency of SCID-repopulating cells derived from umbilical cord blood and granulocyte colony-stimulating factor-mobilized peripheral blood
AU - Pollok, Karen
AU - Hanenberg, Helmut
AU - Williams, David A.
AU - Van der Loo, Johannes C M
AU - Cooper, Ryan J.
AU - Hartwell, Jennifer R.
AU - Miles, Kathryn R.
AU - Breese, Robert
AU - Williams, Emily P.
AU - Montel, Angela
AU - Seshadri, Roopa
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0035684026&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035684026&partnerID=8YFLogxK
U2 - 10.1089/10430340152677430
DO - 10.1089/10430340152677430
M3 - Article
C2 - 11747599
AN - SCOPUS:0035684026
VL - 12
SP - 2095
EP - 2108
JO - Human Gene Therapy
JF - Human Gene Therapy
SN - 1043-0342
IS - 17
ER -