Lentiviral Transfer of the LacZ Gene into Human Endothelial Cells and Human Bone Marrow Mesenchymal Stem Cells
Lentiviral Transfer of the LacZ Gene into Human Endothelial Cells and Human Bone Marrow Mesenchymal Stem Cells
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LacZ 基因慢病毒转移至人内皮细胞和人骨髓间充质干细胞
DOI：
10.3727/000000002783985620
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发表时间：
2002
影响因子：
3.3
通讯作者：
N. Tanaka
中科院分区：
文献类型：
--
作者：
T. Totsugawa;N. Kobayashi;T. Okitsu;H. Noguchi;Takamasa Watanabe;T. Matsumura;M. Maruyama;T. Fujiwara;M. Sakaguchi;N. Tanaka
由于基于人类免疫缺陷病毒1型（HIV-1）的慢病毒载体的吸引人的特征之一是它甚至可以感染非分裂细胞，因此慢病毒介导的基因递送系统作为将基因转移到靶细胞中的创新工具目前受到极大的关注。本研究的目的是探讨慢病毒介导的LacZ基因转染人脐静脉内皮细胞（HUVECs）和人骨髓间充质干细胞（HMSCs）的效果。在本研究中，水泡性口炎病毒G蛋白（VSV-G）的假型慢病毒载体编码的E。利用三质粒系统在293T细胞中构建了带有核定位信号（NLS）标签的LacZ基因。使所得慢病毒载体LtV-NLS/LacZ感染HUVEC和HMSC。大约70%的HUVECs呈LacZ表达阳性，50%的HMSCs显示LacZ活性。在两种细胞中，早期和晚期传代阶段之间的转导功效没有显著差异。LtV-NLS/LacZ转导的HUVEC显示内皮标志物的基因表达，包括血管内皮生长因子（VEGF）受体的CD34和flt-1和KDR/flk-1，并且在Matrigel测定中具有与原代培养的HUVEC一样有效的血管生成潜力。这些发现提供了证据，慢病毒载体是有效的工具，基因转移和表达在人类内皮细胞和干细胞，可能是有用的组织工程。
Because one of the attractive characteristics of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors is that it can infect even nondividing cells, a lentivirus-mediated gene delivery system is currently being paid a great deal of attention as an innovative tool for gene transfer into target cells. The purpose of the work was to investigate the efficacy of lentiviral transfer of the LacZ gene into human umbilical vein endothelial cells (HUVECs) and human bone marrow mesenchymal stem cells (HMSCs) in vitro. For the present study, a vesicular stomatitis virus G-protein (VSV-G)-pseudotyped lentiviral vector encoding the E. coli LacZ gene tagged with nuclear localization signal (NLS) was generated in 293T cells by means of the three-plasmid system. The resulting lentiviral vector, LtV-NLS/LacZ, was allowed to infect HUVECs and HMSCs. Approximately 70% of HUVECs were positive for LacZ expression and 50% of HMSCs showed LacZ activity. There was no significant difference in transduction efficacy between early and late-passage phases in both cells. LtV-NLS/LacZ-transduced HUVECs showed gene expression of endothelial markers including CD34 and flt-1 and KDR/flk-1 of vascular endothelial growth factor (VEGF) receptors and had angiogenic potential as efficiently as primarily cultured HUVECs in a Matrigel assay. These findings provide evidence that lentiviral vectors are efficient tools for gene transfer and expression in human endothelial cells and stem cells that could be useful for tissue engineering.
DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
DOI：
10.1073/pnas.91.10.4407
发表时间：
1994-05-10
影响因子：
11.1
作者：
YANG, YP;NUNES, FA;WILSON, JM
通讯作者：
WILSON, JM
DOI：
10.1073/pnas.90.17.8033
发表时间：
1993-09-01
影响因子：
11.1
作者：
BURNS, JC;FRIEDMANN, T;YEE, JK
通讯作者：
YEE, JK