Gene transfer into human umbilical cord blood-derived CD34+ cells by particle-mediated gene transfer.
Delivery of genes into hematopoietic progenitor cells offers an attractive means for the introduction of corrective or protective genes into cells of both the myeloid and lymphoid lineage. Previously, investigators have often used murine retroviral vectors for gene delivery which require cells to be cycling for efficient delivery. We describe a nonviral method of gene delivery using particle-mediated gene transfer to obviate many disadvantages of viral vectors related to safety, production costs and the need for cell cycle proliferation. Using a CMV-CAT reporter plasmid, we show transfection of highly purified CD34+ cells isolated from umbilical cord blood. Effective gene transfer was shown in unstimulated and in growth-stimulated cells. Following transfection with a neomycin resistance gene, differentiation into cells of the myeloid lineage was observed, assayed by CFU-GM in the presence of G-418. Both unstimulated and stimulated cells gave rise to CFU-GM in the presence of G-418, indicating that stable expression of the neomycin resistance gene was maintained in early progenitors. These results demonstrate that particle-mediated gene transfer into human hematopoietic cells from umbilical cord blood can be achieved without affecting their CFU-GM differentiation potential. This gene transfer method offers an alternative approach to gene therapy studies involving human hematopoietic progenitor cells.
Verma
S
S
Howard Hughes Medical Institute, University of Michigan Medical Center, Department of Internal Medicine, Ann Arbor 48109-0650, USA.
Woffendin
C
C
Bahner
I
I
Ranga
U
U
Xu
L
L
Yang
Z Y
ZY
King
S R
SR
Kohn
D B
DB
Nabel
G J
GJ
D000900:Anti-Bacterial Agents; D018952:Antigens, CD34 / Q000235:genetics* / Q000302:isolation & purification; D015139:Blotting, Southern; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018920:Coculture Techniques; D003587:Cytomegalovirus / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D005312:Fetal Blood / Q000166:cytology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D017979:Gold Colloid; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D009355:Neomycin; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
D000900:Anti-Bacterial Agents; D018952:Antigens, CD34; D005839:Gentamicins; D017979:Gold Colloid; C010680:antibiotic G 418; D015500:Chloramphenicol O-Acetyltransferase; D009355:Neomycin