Constructing tissue-engineered adipose with the combination of gene-transfected human umbilical cord mesenchymal stem cells and silk fibroin scaffold

doi:10.3969/j.issn.2095-4344.2013.14.005

Abstract

Abstract:

BACKGROUND: The method of construction of tissue-engineered adipose is imperfect at present. Although the tissue-engineered adipose can be constructed, the efficacy is not satisfactory.
OBJECTIVE: To study the new method of construction of tissue-engineered adipose, in order to observe capacity of human umbilical cord mesenchymal stem cells transferred with recombinant insulin gene lentiviral vector combined with silk fibroin scaffold in the construction of tissue engineering adipose in Wistar rats.
METHODS: Human umbilical cord mesenchymal stem cells were separated and cultured, and then transfected with recombinant insulin gene lentiviral vector (transfected group) by the best multiplicity of infection =10. The nontransfected human umbilical cord mesenchymal stem cells were regarded as control group. The human umbilical cord mesenchymal stem cells in the transfected group and the control group were seeded onto the silk fibroin scaffold and implanted into the subcutaneous layer of Wistar rats. At 12 weeks after implantation, the transplants were taken, and then identified with fluorescence in situ hybridization and observed with histomorphology and scanning electron microscopy.
RESULTS AND CONCLUSION: Oil red O staining showed the transplants in two groups were positive, suggesting that the transplants were synthesized in adipose tissue, and the number of fat-like cells in the transfected group was significantly higher than that in the control group (P < 0.01). Hematoxylin-eosin staining showed significant angiogenesis appeared in or around the new formed tissue, the structure of which was similar to natural adipose tissue. Silk fibroin scaffold in the transfected group was degraded significantly, the number of new vessels in the transfected group was more than that in the control group, and inflammatory cell infiltration in the transfected group was significantly less than that in the control group. Scanning electron microscopy results showed that fat-like cells in the trasnfected group congregated and the structure was similar to that of the normal adipose tissue; the fat-like cells in the control group scattered in the pore of the scaffold. Insulin gene could obviously promote human umbilical cord mesenchymal stem cells to differentiate into adipose; human umbilical cord mesenchymal stem cells transferred with recombinant human insulin gene lentiviral vector composite with silk fibroin scaffolds can construct tissue-engineered adipose in the Wistar rats, and its structure is similar to natural adipose.

Key words: stem cells, umbilical cord blood stem cells, tissue-engineered adipose, human umbilical cord mesenchymal stem cells, silk fibroin, scaffold, insulin, gene transfection, lentiviral vector, transplantation, Wistar rats, in vivo experiment, National Natural Science Foundation of China, stem cell photographs-containing paper

CLC Number:

R394.2

Liu Yi, Tang Jun, Li Shi-long. Constructing tissue-engineered adipose with the combination of gene-transfected human umbilical cord mesenchymal stem cells and silk fibroin scaffold[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(14): 2501-2508.

Figures/Tables 6

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