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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2003.1
- 수록면
- 412 - 420 (9page)
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We report the transduction of human VEGF165 the transduced myoblasts for transduction and expression efficiency. Human myoblasts were as-sessed by imunostaining for desmin expression. A replication incompetent adenoviral vector cary-ing human VEGF165 was constructed and used for transduction of myoblasts. Imunostaining of trans-duced myoblasts was used to determine trans-duction efficiency. Expression eficiency was con-firmed by imunobloting, ELISA and reverse transcription (RT)-PCR analysis using human-VEGF165 specific primers (5'-3' = 5'ATGACTCT-CTTGTCACA3. Biological activity of the secreted VEGF165 was determined by human umbilical vein endothelial cell proliferation and [H3] thymidine incorporation assays. Human myoblast preparation was＞95% pure with 99% viability after transduc-tion. Imunostaining showed＞95% VEGF165 posi-tive myoblasts. Western bloting and ELISA re-vealed high VEGF165 expression in the transduced myoblasts. Maximum transduction efficiency was at 1:1,000 ratio on three consecutive days. Con-centration of VEGF165 released in the culture med-ium peaked (373 ng/ml) at 8 days post-trans-duction. Cell proliferation asay on human um-bilical vein endothelial cells using supernatant from VEGF165 transduced myoblasts revealed ex-tensive proliferation of cells which was supres-sed in the presence of anti-human VEGF165 anti-body in culture medium and was further confirmed by thymidine incorporation assay. The untrans-duced myoblasts secreted VEGF165 in vitro (30 50 pg/ml) that is enhanced many folds (373 ng/ml) in VEGF165 transduced myoblast as deter-mined by ELISA. These studies suggest that human myoblast are potential cariers of human VEGF165 to achieve concurrent angiomyogenesis for cardiac repair.
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