Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1992-2-18
pubmed:abstractText
Retroviral infection of bone marrow cells in long-term marrow cultures (LTMCs) offers several theoretical advantages over other methods for gene transfer into hematopoietic stem cells. To investigate the feasibility of this approach in a large animal model system, we subjected LTMCs from nine dogs to multiple infections with retrovirus containing the neomycin phosphotransferase gene (neo) during 21 days of culture. Feeder layers, cocultivation, polycations, and selection were not used. The in vitro gene transfer efficiency was 70% as determined by polymerase chain reaction amplification of neo sequences in colony-forming unit granulocyte-macrophage (CFU-GM) obtained from day-21 LTMCs. Day-21 LTMC cells were infused into autologous recipients with (four dogs) and without (three dogs) marrow-ablative conditioning. At 3 months posttransplant, up to 10% of marrow cells contained the neo gene. This percentage declined to 0.1% to 1% at 10 to 21 months posttransplant. Neo was also detected in individual CFU-GM, burst-forming unit-erythroid (BFU-E), and CFU-Mix progenitors derived from marrow up to 21 months postinfusion and in cultures of peripheral blood-derived T cells up to 19 months postinfusion. There was no difference in the percentage of neo-marked cells present when dogs that received marrow ablative conditioning were compared with dogs receiving no conditioning. Detection of neo-marked marrow cells almost 2 years after autologous transplantation in a large mammalian species shows that retroviral infection of marrow cells in LTMCs is a potentially nontoxic and efficient protocol for gene transfer. Further, our results suggest that marrow conditioning and in vivo selection pressure to retain transplanted cells may not be absolute requirements for the retention of genetically marked cells in vivo.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
AIM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0006-4971
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
79
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
356-64
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:1309669-Animals, pubmed-meshheading:1309669-Base Sequence, pubmed-meshheading:1309669-Bone Marrow, pubmed-meshheading:1309669-Bone Marrow Cells, pubmed-meshheading:1309669-Bone Marrow Transplantation, pubmed-meshheading:1309669-Cells, Cultured, pubmed-meshheading:1309669-DNA, pubmed-meshheading:1309669-Dogs, pubmed-meshheading:1309669-Genetic Markers, pubmed-meshheading:1309669-Genetic Vectors, pubmed-meshheading:1309669-Granulocytes, pubmed-meshheading:1309669-Hematopoietic Stem Cells, pubmed-meshheading:1309669-Kanamycin Kinase, pubmed-meshheading:1309669-Macrophages, pubmed-meshheading:1309669-Molecular Sequence Data, pubmed-meshheading:1309669-Phosphotransferases, pubmed-meshheading:1309669-Polymerase Chain Reaction, pubmed-meshheading:1309669-Retroviridae, pubmed-meshheading:1309669-Transfection
pubmed:year
1992
pubmed:articleTitle
Autologous transplantation of canine long-term marrow culture cells genetically marked by retroviral vectors.
pubmed:affiliation
University of Toronto Hospitals' Cancer Cytogenetics Program, Toronto General Hospital, Ontario, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't