Korean J Med.  2006 Mar;70(3):288-297.

hVEGF121 transfection and animal model for in vivo angiogenesis

• Park JI
• Choi YJ
• Choi JH
• Byun J
• Kim DK

Abstract

BACKGROUND: This study was designed to prove the superiority of ACP vector containing ITR, and to establish animal model quantifying angiogenesis in vivo.
METHODS
hVEGF121, therapeutic gene, was inserted to various vectors (pcDNA3.1, pcDNA 3.2, pActin, pDesm, pACP vector), and these vectors were transfected to various cells using FuGENE6. We cultured for 48hrs, and then quantified amounts of hVEGF121 of supernatants by ELISA. The long-term transfection was assessed for 14 days. Optimal condition of transfection was evaluated by change of the ratio of DNA to FuGENE6, amount of DNA, and confluence of cells. ACP-hVEGF121 was transfected to C2C12 and these transfected C2C12 cells were mixed with Matrigel, and then injected to C3H mouse subcutaneously. Seven days later, hemoglobin assay and pathology of Matrigel were reviewed for angiogenesis.
RESULTS
The level of hVEGF121 gene expression using pACP vector was significantly higher than those of others. In 2 weeks culture study, pACP vector showed the highest gene expression and produced VEGF until 2 weeks. The highest gene expression was obtained when the concentration of DNA was 7 microgram, the confluence was up to 80% and the ratio of DNA to FuGENE6 was 1:3. The hemoglobin level in Matrigel of VEGF group was significantly higher than the one of the control group, and active angiogenesis was noted in the VEGF group.
CONCLUSIONS
pACP vector might be an efficient vector for angiogenic gene delivery, and animal model using Matrigel and transfected C2C12 cell could be a useful tool for quantitative angiogenesis assay.

Keyword

ACP vector; Vascular Endothelial Growth Factor; Angiogenesis Assay

MeSH Terms

Animals*
DNA
Enzyme-Linked Immunosorbent Assay
Gene Expression
Mice
Mice, Inbred C3H
Models, Animal*
Pathology
Transfection*
Vascular Endothelial Growth Factor A
DNA
Vascular Endothelial Growth Factor A