|
|
 Previous Article | Table of Contents | Next Article 
Gene transfer into normal human hematopoietic cells using in vitro and in
vivo assays
JE Dick, S Kamel-Reid, B Murdoch and M Doedens
Department of Genetics, Hospital for Sick Children, Toronto, Ontario,
Canada.
The ability to transfer new genetic material into human hematopoietic cells
provides the foundation for characterizing the organization and
developmental program of human hematopoietic stem cells. It also provides a
valuable model in which to test gene transfer and long-term expression in
human hematopoietic cells as a prelude to human gene therapy. At the
present time such studies are limited by the absence of in vivo assays for
human stem cells, although recent descriptions of the engraftment of human
hematopoietic cells in immune-deficient mice may provide the basis for such
an assay. This study focuses on the establishment of conditions required
for high efficiency retrovirus- mediated gene transfer into human
hematopoietic progenitors that can be assayed in vitro in short-term colony
assays and in vivo in immune- deficient mice. Here we report that a 24-hour
preincubation of human bone marrow in 5637-conditioned medium, before
infection, increases gene transfer efficiency into in vitro colony-forming
cells by sixfold; interleukin-6 (IL-6) and leukemia inhibitory factor (LIF)
provide the same magnitude increase as 5637-conditioned medium. In
contrast, incubation in recombinant growth factors IL-1, IL-3, and
granulocyte- macrophage colony-stimulating factor increases gene transfer
efficiency by 1.5- to 3-fold. Furthermore, preselection in high
concentrations of G418 results in a population of cells significantly
enriched for G418- resistant progenitors (up to 100%). These results,
obtained using detailed survival curves based on colony formation in G418,
have been substantiated by directly detecting the neo gene in individual
colonies using the polymerase chain reaction. Using these optimized
protocols, human bone marrow cells were genetically manipulated with a neo
retrovirus vector and transplanted into immune-deficient bg/nu/xid mice. At
1 month and 4 months after the transplant, the hematopoietic tissues of
these animals remained engrafted with genetically manipulated human cells.
More importantly, G418-resistant progenitors that contained the neo gene
were recovered from the bone marrow and spleen of engrafted animals after 4
months. These experiments establish the feasibility of characterizing human
stem cells using the unique retrovirus integration site as a clonal marker,
similar to techniques developed to elucidate the murine stem cell
hierarchy.
Volume 78,
Issue 3,
pp. 624-634,
08/01/1991
Copyright © 1991 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:
|
|
|
|
 
A. Rovira, M. De Angioletti, O. Camacho-Vanegas, D. Liu, V. Rosti, H. F. Gallardo, R. Notaro, M. Sadelain, and L. Luzzatto
Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer
Blood,
December 15, 2000;
96(13):
4111 - 4117.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Hennemann, I.-H. Oh, J. Y. Chuo, C. P. Kalberer, P. D. Schley, S. Rose-John, R. K. Humphries, and C. J. Eaves
Efficient retrovirus-mediated gene transfer to transplantable human bone marrow cells in the absence of fibronectin
Blood,
October 1, 2000;
96(7):
2432 - 2439.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. F. Kelly, J. Vandergriff, A. Nathwani, A. W. Nienhuis, and E. F. Vanin
Highly efficient gene transfer into cord blood nonobese diabetic/severe combined immunodeficiency repopulating cells by oncoretroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein
Blood,
August 15, 2000;
96(4):
1206 - 1214.
[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]
|
|
|
|
|
|
|
S. S. Case, M. A. Price, C. T. Jordan, X. J. Yu, L. Wang, G. Bauer, D. L. Haas, D. Xu, R. Stripecke, L. Naldini, et al.
Stable transduction of quiescent CD34+CD38- human hematopoietic cells by HIV-1-based lentiviral vectors
PNAS,
March 16, 1999;
96(6):
2988 - 2993.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Veena, C. M. Traycoff, D. A. Williams, J. McMahel, S. Rice, K. Cornetta, and E. F. Srour
Delayed Targeting of Cytokine-Nonresponsive Human Bone Marrow CD34+ Cells With Retrovirus-Mediated Gene Transfer Enhances Transduction Efficiency and Long-Term Expression of Transduced Genes
Blood,
May 15, 1998;
91(10):
3693 - 3701.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. L. Greiner, R. A. Hesselton, and L. D. Shultz
SCID Mouse Models of Human Stem Cell Engraftment
Stem Cells,
May 1, 1998;
16(3):
166 - 177.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
J.C.Y. Wang, T. Lapidot, J.D. Cashman, M. Doedens, L. Addy, D.R. Sutherland, R. Nayar, P. Laraya, M. Minden, A. Keating, et al.
High Level Engraftment of NOD/SCID Mice by Primitive Normal and Leukemic Hematopoietic Cells From Patients With Chronic Myeloid Leukemia in Chronic Phase
Blood,
April 1, 1998;
91(7):
2406 - 2414.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. K. Fielding, M. Maurice, F. J. Morling, F.-L. Cosset, and S. J. Russell
Inverse Targeting of Retroviral Vectors: Selective Gene Transfer in a Mixed Population of Hematopoietic and Nonhematopoietic Cells
Blood,
March 1, 1998;
91(5):
1802 - 1809.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. D. Lyman and S. E. W. Jacobsen
c-kit Ligand and Flt3 Ligand: Stem/Progenitor Cell Factors With Overlapping Yet Distinct Activities
Blood,
February 15, 1998;
91(4):
1101 - 1134.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Yurasov, T. R. Kollmann, A. Kim, C. A. Raker, M. Hachamovitch, F. Wong-Staal, and H. Goldstein
Severe Combined Immunodeficiency Mice Engrafted With Human T Cells, B Cells, and Myeloid Cells After Transplantation With Human Fetal Bone Marrow or Liver Cells and Implanted With Human Fetal Thymus: A Model for Studying Human Gene Therapy
Blood,
March 1, 1997;
89(5):
1800 - 1810.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M.K. Brenner, H.E. Heslop, D. Rill, C. Li, T. Nilson, M. Roberts, C. Smith, R. Krance, and C. Rooney
Gene Transfer and Bone Marrow Transplantation
Cold Spring Harb Symp Quant Biol,
January 1, 1994;
59(0):
691 - 697.
[Abstract]
[PDF]
|
|
|
|
|
|
|
C. J. Schmeichel and J. M. Loeb
Human Gene Therapy: A Role for the Primary Care Physician
Arch Fam Med,
September 1, 1992;
1(1):
113 - 120.
[Abstract]
[PDF]
|
|
|
|
|
|
|
T Lapidot, F Pflumio, M Doedens, B Murdoch, D. Williams, and J. Dick
Cytokine stimulation of multilineage hematopoiesis from immature human cells engrafted in SCID mice
Science,
February 28, 1992;
255(5048):
1137 - 1141.
[Abstract]
[PDF]
|
|
|
|
|
