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Transgene expression and differentiation of baculovirus‐transduced human mesenchymal stem cells
Author(s) -
Ho YiChen,
Chung YaoChi,
Hwang ShiawMin,
Wang KueiChun,
Hu YuChen
Publication year - 2005
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.729
Subject(s) - transduction (biophysics) , transgene , biology , mesenchymal stem cell , genetic enhancement , green fluorescent protein , progenitor cell , stem cell , gene delivery , viral vector , microbiology and biotechnology , transfection , cell culture , gene , recombinant dna , genetics , biochemistry
Abstract Background Mesenchymal stem cells (MSCs) have drawn considerable attention as vehicles for cell‐ or gene‐based therapies, yet various problems still exist for current gene delivery vectors. On the other hand, baculovirus has emerged as a novel gene therapy vector, but its transduction of stem cells has not been reported. Methods A recombinant baculovirus expressing the enhanced green fluorescent protein (EGFP) was constructed to transduce human MSCs derived from umbilical cord blood (uMSCs) or bone marrow (bMSCs). Results In this study, we demonstrated for the first time that human uMSCs or bMSCs could be transduced by baculovirus with high efficiencies (up to ≈72.8% and 41.1%, respectively) and significantly elevated transgene (enhanced green fluorescent protein, EGFP) expression upon incubation with unconcentrated virus and phosphate‐buffered saline for 4 h at 25 °C. The transduction efficiency into bMSCs could be further increased to ≈72.2% by lowering the cell density. The improved transgene expression was partly attributed to the enhanced virus uptake upon transduction, as determined by quantitative real‐time polymerase chain reaction (Q‐PCR). MSC growth was not obstructed by baculovirus transduction itself, but was somewhat hampered by EGFP expression. Nonetheless, the baculovirus‐transduced cells remained capable of differentiating into adipogenic lineage. The adipogenic progenitors appeared more permissive to baculovirus transduction than the undifferentiated bMSCs, thus allowing for the maintenance and enhancement of transgene expression by repeated transduction after subculture. Conclusions These findings implicate the potential applications of baculovirus as an alternative vector to genetically modify MSCs for ex vivo gene therapy. Copyright © 2005 John Wiley & Sons, Ltd.