Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
6
|
| pubmed:dateCreated |
1992-2-20
|
| pubmed:abstractText |
Liver-directed gene therapy is being considered in the treatment of inherited metabolic diseases. One approach we are considering is the transplantation of autologous hepatocytes that have been genetically modified with recombinant retroviruses ex vivo. We describe, in this report, techniques for isolating human hepatocytes and efficiently transducing recombinant genes into primary cultures. Hepatocytes were isolated from tissue of four different donors, plated in primary culture, and exposed to recombinant retroviruses expressing either the LacZ reporter gene or the cDNA for rabbit LDL receptor. The efficiency of gene transfer under optimal conditions, as determined by Southern blot analysis, varied from a maximum of one proviral copy per cell to a minimum of 0.1 proviral copy per cell. Cytochemical assays were used to detect expression of the recombinant derived proteins, E. coli beta-galactosidase and rabbit LDL receptor. Hepatocytes transduced with the LDL receptor gene expressed levels of receptor protein that exceeded the normal endogenous levels. The ability to isolate and genetically modify human hepatocytes, as described in this report, is an important step towards the development of liver-directed gene therapies in humans.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0740-7750
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
17
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
601-7
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:1767337-Adolescent,
pubmed-meshheading:1767337-Adult,
pubmed-meshheading:1767337-Blotting, Southern,
pubmed-meshheading:1767337-Cell Separation,
pubmed-meshheading:1767337-Cells, Cultured,
pubmed-meshheading:1767337-Child, Preschool,
pubmed-meshheading:1767337-DNA, Recombinant,
pubmed-meshheading:1767337-Female,
pubmed-meshheading:1767337-Gene Therapy,
pubmed-meshheading:1767337-Genetic Vectors,
pubmed-meshheading:1767337-Histocytochemistry,
pubmed-meshheading:1767337-Humans,
pubmed-meshheading:1767337-Infant,
pubmed-meshheading:1767337-Lac Operon,
pubmed-meshheading:1767337-Liver,
pubmed-meshheading:1767337-Male,
pubmed-meshheading:1767337-Nucleic Acid Hybridization,
pubmed-meshheading:1767337-Recombinant Fusion Proteins,
pubmed-meshheading:1767337-Retroviridae,
pubmed-meshheading:1767337-Transfection
|
| pubmed:year |
1991
|
| pubmed:articleTitle |
Towards liver-directed gene therapy: retrovirus-mediated gene transfer into human hepatocytes.
|
| pubmed:affiliation |
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor 48109.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|