Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (6): 999-1003.doi: 10.3969/j.issn.2095-4344.2013.06.009

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Adipogenic differentiation of human umbilical cord mesenchymal stem cells transfected by insulin gene-modified adenovirus vector

Li Shi-long1, Liu Yi2, Hui Ling3   

  1. 1 The Second Clinical Medical College of Lanzhou University, Lanzhou 730050, Gansu Province, China
    2 Center of Military Burns and Plastic Surgery, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
    3 Gansu Provincial Key Laboratory of Stem Cells and Gene Drugs, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Received:2012-05-03 Revised:2012-06-15 Online:2013-02-05 Published:2013-02-05
  • Contact: Liu Yi, Chief physician, Professor, Center of Military Burns and Plastic Surgery, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou 730050, Gansu Province, China 1iuzhih20002003@yahoo.com.cn
  • About author:Li Shi-long, the Second Clinical Medical College of Lanzhou University, Lanzhou 730050, Gansu Province, China lsllwxkp@126.com

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Abstract:

BACKGROUND: Insulin is an important component of adipogenic inducers. Insulin has the half-life, which can continuously inactivate and degrade with the in vivo metabolism, and the cells and the materials implanted in the body cannot replace the culture medium to achieve the purpose like in in vitro environment. Transfection of insulin gene can make the transfected stem cells secret the insulin stably which can promote the adipogenic differentiation.
OBJECTIVE: To investigate the adipogenic differentiation potential of human umbilical cord mesenchymal stem cells transfected by insulin gene-modified adenovirus vector.
METHODS: Passage 4 human umbilical cord mesenchymal stem cells were collected and transfected with adenovirus vector with the multiplicity of infection of 20. The experiment was divided into four groups. Passage 4 human umbilical cord mesenchymal stem cells were included in the control group. Passage 4 human umbilical cord mesenchymal stem cells transfected with adenovirus vector were included in experimental group 1, passage 4 human umbilical cord mesenchymal stem cells + adipogenic liquid were used in the experimental group 2, and passage 4 human umbilical cord mesenchymal stem cells transfected with adenovirus vector + adipogenic liquid were used in experimental group 3.
RESULTS AND CONCLUSION: After transfected for 48 hours, weak fluorescence in human umbilical cord mesenchymal stem cells was observed in experimental groups1 and 3 through the fluorescence microscope and strong fluorescence appeared after transfected for 72 hours After cultured with adipose-inducing culture medium for 14 days, the cells in the experimental groups 1, 2, 3 were red under microscope after oil red O staining, no lipid droplets were observed in the control group; the lipid droplets in the experimental group 3 were larger and more than those in the experimental group 2. Quantitative analysis showed the positive area of oil red O staining and the absorbance value in the experimental group 3 were greater than those in the experimental group 2 (P < 0.05). The results show that the human umbilical cord mesenchymal stem cells transfected by insulin gene-modified adenovirus vector can promote cell adipogenic differentiation.

Key words: stem cells, umbilical cord/umbilical cord blood stem cells, human insulin gene, mesenchymal stem cells, adenovirus vector, gene transfection, cell differentiation, other grants-supported paper, stem cell photographs-containing paper

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