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Joong Il Park, Yu Jeong Choi, Jin Ho Choi, Jong Hoe Byun, Duk Kyung Kim |
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이화여자대학교 의과대학 내과학 교실, 의과학 연구소 |
| 원저 : C2C12 세포에서의 pACP 벡터에 의한 hVEGF121 전달 및 in vivo 혈관신생 정량 모델의 확립 (Original Articles : hVEGF121 transfection and animal model for in vivo angiogenesis) |
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박중일, 최유정, 최진호, 변종회, 김덕경 |
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| 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 μg, 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.(Korean J Med 70:288-297, 2006)
Key Words : ACP vector, Vascular Endothelial Growth Factor, Angiogenesis Assay |
| Key Words:
ACP vector, Vascular Endothelial Growth Factor, Angiogenesis Assay |
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