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[Enhanced green fluorescence protein reporter gene labeling of mesenchymal stem cells mediated by lentivirus].

Authors :
Li D
Zhu KS
Zhou B
Wang J
Yan RH
Li ZR
Meng XC
Huang MS
Jiang ZB
Shan H
Source :
Zhonghua yi xue za zhi [Zhonghua Yi Xue Za Zhi] 2010 May 18; Vol. 90 (19), pp. 1357-61.
Publication Year :
2010

Abstract

Objective: To explore the effect of enhanced green fluorescence protein (EGFP) labeling mediated by lentivirus on the biophysical properties of mesenchymal stem cells (MSC), and whether the EGFP gene expression is permanent and stable.<br />Methods: MSC were infected with EGFP lentivirus at different virus multiplicity of infection (MOI). EGFP positive rate was measured with fluorescent-activated cell scanning (FACS) analysis, and EGFP expression in MSC was investigated under a fluorescence microscope. Cell viability, proliferation, apoptosis and cell cycle were detected with trypan blue stain, MTT colorimetric assay, Hoechst stain and FACS analysis respectively. To evaluate the stability of EGFP expression, EGFP lentivirus infected MSC were harvested after cultured continuously in vitro for 2, 4, 8 or 16 weeks, and EGFP positive rate and fluorescence strength were detected with FACS analysis.<br />Results: After infected with EGFP lentivirus (MOI = 20) for 96 h, EGFP positive rate of MSC was 97.39% +/- 0.68%. Cell viability, proliferation, apoptosis and cell cycle of MSC infected with EGFP lentivirus were unaffected, as compared with control MSC (P > 0.05). When cultured in vitro continuously for 2, 4, 8 or 16 weeks, EGFP positive rates of EGFP-MSC were 97.50% +/- 0.54%, 97.32% +/- 0.51%, 97.39% +/- 0.11%, and 97.48% +/- 0.13% respectively, while EGFP fluorescence strength were 440 +/- 13, 445 +/- 12, 458 +/- 13 and 456 +/- 16 respectively. Both EGFP positive rate and fluorescence strength kept in a stable level.<br />Conclusion: EGFP lentivirus can efficiently label MSC and has no significant effect on the biophysical properties of MSC. EGFP gene expression in MSC is permanent and stable. EGFP-MSC can be used for further cell tracing research.

Details

Language :
Chinese
ISSN :
0376-2491
Volume :
90
Issue :
19
Database :
MEDLINE
Journal :
Zhonghua yi xue za zhi
Publication Type :
Academic Journal
Accession number :
20646588