| Baculovirus transduction of rat articular chondrocytes: roles of cell cycle. | |
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MedLine Citation:
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PMID: 17167815 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: We have previously demonstrated highly efficient baculovirus transduction of primary rat articular chondrocytes, thus implicating the possible applications of baculovirus in gene-based cartilage tissue engineering. However, baculovirus-mediated gene expression in the chondrocytes is transient. METHODS: In this study, we attempted to prolong the expression by supertransduction, but uncovered that after long-term culture the chondrocytes became more refractory to baculovirus transduction. Therefore, the correlation between baculovirus-mediated enhanced green fluorescent protein (EGFP) expression and cell cycle was investigated by comparing the cycling chondrocytes and chondrocytes rich in quiescent cells, in terms of EGFP expression, virus uptake, cell cycle distribution, nuclear import and methylation of viral DNA. RESULTS: We demonstrated, for the first time, that baculovirus-mediated transduction of chondrocytes is correlated with the cell cycle. The chondrocytes predominantly in G2/M phase were approximately twice as efficient in EGFP expression as the cycling cells, while the cells in S and G1 phases expressed EGFP as efficiently as the cycling cells. Notably, the chondrocyte populations rich in quiescent cells resulted in efficient virus uptake, but less effective nuclear transport of baculoviral DNA and higher degree of methylation, and hence poorer transgene expression. CONCLUSIONS: These findings unravel the practical limitations when employing baculovirus in cartilage tissue engineering. The implications and possible solutions are discussed. |
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Authors:
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Hsiao-Ping Lee; Yen-Lin Chen; Heng-Chun Shen; Wen-Hsin Lo; Yu-Chen Hu |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The journal of gene medicine Volume: 9 ISSN: 1099-498X ISO Abbreviation: J Gene Med Publication Date: 2007 Jan |
Date Detail:
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Created Date: 2007-01-08 Completed Date: 2007-03-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9815764 Medline TA: J Gene Med Country: England |
Other Details:
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Languages: eng Pagination: 33-43 Citation Subset: IM |
Copyright Information:
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Copyright 2006 John Wiley & Sons, Ltd. |
Affiliation:
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Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan 300. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Active Transport, Cell Nucleus Animals Baculoviridae / genetics* Cartilage, Articular / cytology* Cell Cycle* Cells, Cultured Chondrocytes / cytology, physiology* DNA Methylation Gene Expression Green Fluorescent Proteins / genetics, metabolism Rats Rats, Wistar Tissue Engineering Transduction, Genetic* Transgenes |
| Chemical | |
Reg. No./Substance:
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147336-22-9/Green Fluorescent Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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