【摘 要】
：
目的 探讨重组腺相关病毒载体能否有效地转染脂肪组织来源基质干细胞及(ADSC)其影响,同时为组织工程寻找一种理想的病毒裁体及细胞示踪标记方法.方法 采用抽脂法分离培养ADSC
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目的 探讨重组腺相关病毒载体能否有效地转染脂肪组织来源基质干细胞及(ADSC)其影响,同时为组织工程寻找一种理想的病毒裁体及细胞示踪标记方法.方法 采用抽脂法分离培养ADSC,诱导培养液诱导细胞成脂分化,行细胞形态学检查,细胞表面抗原鉴定,油红O染色,评价细胞成脂情况.在此基础上转粢Ad-EGFP,观察细胞形态学改变及荧光表达的时间与强度,计算转粢效率,确定转染的理想感染复数(MOI)值.同时通过活力和增殖实验进行Ad-EGFP体外转染对ADSC的影响研究.结果 细胞扩增迅速,形态良好,经诱导后呈现明显的成脂改变.实验确定Ad转染细胞的较佳MOI值为50,以此值转染Ad-EGFP后细胞荧光持续高效稳定表达,并可传至子代,可以满足示踪标记的需要.Ad转染效率高,对细胞活性影响小.结论 ADSC的培养和分化能满足脂肪组织工程的要求;Ad长期稳定表达目的 基因,是一种理想的组织工程病毒裁体;基因转染方法高效稳定表达EGFP,可作为种子细胞良好的示踪标记方法.
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