Tumor necrosis factor-related apoptosis-inducing ligand modified human amniotic fluid-derived mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.23.013

Abstract

Abstract:

BACKGROUND: The new gene therapy with mesenchymal stem cells as carriers has wide application value.
OBJECTIVE: To import tumor necrosis factor-related apoptosis-inducing ligand into human amniotic fluid-derived mesenchymal stem cells using lentiviral vector, in order to obtain human amniotic fluid-derived mesenchymal stem cells that stably express tumor necrosis factor-related apoptosis-inducing ligand.
METHODS: The lentiviral expression vector pLVpuro/EF1α-tumor necrosis factor-related apoptosis-inducing ligand was constructed by multisite Gateway technique. The expression vector and the lentiviral packaging plasmid were co-transfected with 293FT cells to obtain the lentiviral plasmid carrying tumor necrosis factor-related apoptosis-inducing ligand. The human amniotic fluid-derived mesenchymal stem cells were transfected with recombinant lentivirus particles, and then the human amniotic fluid-derived mesenchymal stem cells that stably express target gene-tumor necrosis factor-related apoptosis-inducing ligand were obtained by using antibiotic screening methods. The stability of human amniotic fluid-derived mesenchymal stem cells was detected.
RESULTS AND CONCLUSION: The enzyme-linked immunosorbent assay and western blot results indicated that the tumor necrosis factor-related apoptosis-inducing ligand protein could be highly expressed in the transfected human amniotic fluid-derived mesenchymal stem cells, and the concentration of tumor necrosis factor-related apoptosis-inducing ligand protein could reach 72 μg/L. These results suggest that the tumor necrosis factor-related apoptosis-inducing ligand modified human amniotic fluid-derived mesenchymal stem cells are established successfully.

Key words: stem cells, stem cell culture and differentiation, human amniotic fluid-derived mesenchymal stem cells, tumor necrosis factor-related apoptosis-inducing ligand, gene modification, lentivirus vector, multi-site, stable expression, target gene, 293FT cell, tissue engineering, National Natural Science Foundation of China, stem cell photographs-containing paper

CLC Number:

R394.2

Du Jing-chun, Zhu Rui, Fan Ting-ting, Wang Peng-kun, Lin Yong-ping, Xu Xia. Tumor necrosis factor-related apoptosis-inducing ligand modified human amniotic fluid-derived mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(23): 4272-4278.

Figures/Tables 7

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