|
|
 Previous Article | Table of Contents | Next Article 
Growth factors and stromal support generate very efficient retroviral
transduction of peripheral blood CD34+ cells from Gaucher patients
LC Xu, S Kluepfel-Stahl, M Blanco, R Schiffmann, C Dunbar and S Karlsson
Molecular and Medical Genetics Section, National Institutes of Neurological
Disorders and Stroke (NINDS), Bethesda, MD 20892, USA.
We have achieved high-efficiency gene transfer into nonmobilized peripheral
blood (PB) CD34+ cells from patients with Gaucher's disease using a
clinically acceptable retroviral supernatant transduction protocol. In our
studies, bone marrow (BM) and PB CD34+ cells were transduced using a high
titer (10(8) particles/mL) retroviral supernatant once a day for 4
consecutive days in the presence of interleukin-3 (IL-3), IL-6, and stem
cell factor (SCF), with or without an irradiated allogeneic BM stromal
layer. The growth factors alone resulted in 29% +/- 10% gene transfer of PB
CD34+ clonogenic cells in contrast with 71% +/- 17% gene transfer
efficiency using stroma with the growth factors; a 2.5-fold increase. The
increase in gene transfer efficiency was less prominent when BM CD34+ cells
were used (40% +/- 16% without and 57% +/- 8% with stroma, a 1.5-fold
increase). The overall transduction efficiency of both PB and BM CD34+
cells was lower when the cells were transduced over a stromal cell layer
without added growth factors. The combination of IL-3, IL-6, and SCF with
stroma transduced 75% of primitive long-term culture initiating cells (PB
LTC- ICs) in comparison with 34% of LTC-ICs when IL-3, IL-6, and SCF were
used without stromal support. Using this clinically acceptable
supernatant/cytokines/stroma transduction protocol, correction of the
glucocerebrosidase (GC) deficiency in the progeny cells of PBLTC-ICs from
Gaucher's-disease patients has been accomplished. Efficient transduction of
the PB CD34+ cells using this transduction protocol may allow repeated
delivery of "GC-corrected" hematopoietic stem and progenitor cells to
Gaucher's-disease patients.
Volume 86,
Issue 1,
pp. 141-146,
07/01/1995
Copyright © 1995 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
B. Schiedlmeier, K. Kuhlcke, H. G. Eckert, C. Baum, W. J. Zeller, and S. Fruehauf
Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice
Blood,
February 15, 2000;
95(4):
1237 - 1248.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. Ramsfjell, D. Bryder, H. Bjorgvinsdottir, S. Kornfalt, L. Nilsson, O. J. Borge, and S. E.W. Jacobsen
Distinct Requirements for Optimal Growth and In Vitro Expansion of Human CD34+CD38- Bone Marrow Long-Term Culture-Initiating Cells (LTC-IC), Extended LTC-IC, and Murine In Vivo Long-Term Reconstituting Stem Cells
Blood,
December 15, 1999;
94(12):
4093 - 4102.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Hibino, K. Tani, K. Ikebuchi, M.-S. Wu, H. Sugiyama, Y. Nakazaki, T. Tanabe, S. Takahashi, A. Tojo, S. Suzuki, et al.
The Common Marmoset as a Target Preclinical Primate Model for Cytokine and Gene Therapy Studies
Blood,
May 1, 1999;
93(9):
2839 - 2848.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. I. Rebel, M. Tanaka, J.-S. Lee, S. Hartnett, M. Pulsipher, D. G. Nathan, R. C. Mulligan, and C. A. Sieff
One-Day Ex Vivo Culture Allows Effective Gene Transfer Into Human Nonobese Diabetic/Severe Combined Immune-Deficient Repopulating Cells Using High-Titer Vesicular Stomatitis Virus G Protein Pseudotyped Retrovirus
Blood,
April 1, 1999;
93(7):
2217 - 2224.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Lutzko, S. Kruth, A. C.G. Abrams-Ogg, K. Lau, L. Li, B. R. Clark, C. Ruedy, S. Nanji, R. Foster, D. Kohn, et al.
Genetically Corrected Autologous Stem Cells Engraft, But Host Immune Responses Limit Their Utility in Canine alpha -L-iduronidase Deficiency
Blood,
March 15, 1999;
93(6):
1895 - 1905.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M.A. Dao, K. Hashino, I. Kato, and J.A. Nolta
Adhesion to Fibronectin Maintains Regenerative Capacity During Ex Vivo Culture and Transduction of Human Hematopoietic Stem and Progenitor Cells
Blood,
December 15, 1998;
92(12):
4612 - 4621.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Povey, N. Weeratunge, C. Marden, A. Sehgal, A. Thrasher, and C. Casimir
Enhanced Retroviral Transduction of 5-Fluorouracil-Resistant Human Bone Marrow (Stem) Cells Using a Genetically Modified Packaging Cell Line
Blood,
December 1, 1998;
92(11):
4080 - 4089.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. L. Maria, C. D. Medina, K. B.N. Hoang, and M. Ian Phillips
Topical Review: Gene Therapy for Neurologic Disease: Benchtop Discoveries to Bedside Applications. 2. The Bedside
J Child Neurol,
February 1, 1997;
12(2):
77 - 84.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. A. Dao, C. H. Hannum, D. B. Kohn, and J. A. Nolta
FLT3 Ligand Preserves the Ability of Human CD34+ Progenitors to Sustain Long-Term Hematopoiesis in Immune-Deficient Mice After Ex Vivo Retroviral-Mediated Transduction
Blood,
January 15, 1997;
89(2):
446 - 456.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
