Ewha Med J.  2002 Mar;25(1):11-16. 10.12771/emj.2002.25.1.11.

The Efficiency of the Vesicular Stomatis G Envelope in Transduction into Hematopoietic Stem Cell

• Park JY
• Yoo ES

• KMID: 2052775
• DOI: http://doi.org/10.12771/emj.2002.25.1.11

Abstract

OBJECTIVES
Stable gene transfer to human pluripotent hematopoietic stem cells is an attractive strategy for the curative treatment of many genetic hematologic disorders. In clinical tral, the level of gene transfer to this cell population have generally been low. In this study we have evaluated the efficiency of gene transfer to human umbilical cord blood(UCB) CD34+cells using vesicular stomatitis virus glycoprotein G(VSV-G) pseudotyped HIV-1 vector. METHOD: High titers of replication-defective VSV-G pseudotyped HIV-1 based vector encoding the enhanced yellow fluorescent protein were produced by transient transfection. Human CD34+cells purified from UCB were incubated with pseudotyped HIV supernatants for 24-48 hours. The transduction efficiency were measured by marker gene expression under the microscopy and flow cytometry.
RESULTS
Transduction rates into CD34+ were low at 0 and 24 hours, reflecting 4.7+/-2.4% at 24 hours, they were increased to 5.7+/-2.7% at 48 hours.
CONCLUSION
We demonstrate efficient transduction of purified human UCB CD34+ by HIV vectors pseudotyped with VSV-G. The results extend the lentiviral vector to clinical gene therapy using human pluripotent hematopoietic stem cells.

Keyword

CD34+; VSV-G; HIV-1 based vector; Lentiviral transduction; Gene therapy

MeSH Terms

Flow Cytometry
Gene Expression
Genetic Therapy
Glycoproteins
Hematopoietic Stem Cells*
HIV
HIV-1
Humans
Microscopy
Transfection
Umbilical Cord
Vesicular Stomatitis
Glycoproteins