Transfection of human umbilical vein endothelial cells with lentivirus containing enhanced green fluorescent protein

doi:10.3969/j.issn.2095-4344.2014.15.010

Abstract

Abstract:

BACKGROUND: Human umbilical vein endothelial cells transfected with lentivirus containing enhanced green fluorescent protein can be easily traced. The optimal multiplicity of infection and time for producing strong fluorescence intensity can lay the foundation of tracing human umbilical vein endothelial cells in animal models.
OBJECTIVE: To observe expression of lentivirus containing enhanced green fluorescent protein in human umbilical vein endothelial cells, and thereby to find a stable method to label human umbilical vein endothelial cells.
METHODS: Using 0.1% collagenase perfusion digestion, we isolated human umbilical vein endothelial cells, which then were placed into a culture medium containing 20% fetal bovine serum and endothelial cell growth factor and observed under an inverted microscope. Following digestion, centrifugation and suspension, the cells were counted and divided into four groups, 5.0×10⁵ cells in each group. After cells were seededonto 24-well plates, 10 μL serum-free Dulbecco’s modified Eagle’s medium was added into the blank group, and lentiviruses containing enhanced green fluorescent protein were added into another three groups for cell transfection respectively at multiplicities of infection of 2, 3, 4. There were three dishes in each group.
RESULTS AND CONCLUSION: After cultured for 5-7 days, isolated cells grew into a single layer and exhibited a cobblestone-like arrangement under a light microscope. In addition, factor VIII related antigen test was positive. A green fluorescence was visible at 24 hours of transfection, and peaked at 72 hours. Transfection efficiency was in a linear growth with the multiplicity of infection. Up to the 21st day of transfection, the green fluorescence was still visible. After 0, 7, 14, 21 days of transfection, the number of human umbilical vein endothelial cells showed no difference between the transfection group with the multiplicity of infection=3 and blank group, suggesting the proliferative ability of cells has no changes after transfection with lentivirus containing enhanced green fluorescent protein. These findings indicate that the lentivirus containing enhanced green fluorescent protein can highly transfect human umbilical vein endothelial cells, and green fluorescent protein can sustainably express for 21 days but cannot impact the cell proliferation.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: lentivirus, green fluorescent proteins, umbilical veins, endothelial cells, transfection

CLC Number:

R318

Ding Qiang, Yang Bo, Wang Le, Yin Biao, Tang Long, Zhang Bo. Transfection of human umbilical vein endothelial cells with lentivirus containing enhanced green fluorescent protein[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(15): 2351-2356.

Figures/Tables 3

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