外源基因在低温保存后同种带瓣大动脉中的表达

Gene transfer to homograft conduit after cold preservation

目的 观察外源性基因在低温保存不同时间后的同种带瓣大动脉中的表达。方法 将 30只幼兔的同种带瓣大动脉分为 5组 ,分别在 4℃营养液中 (M 199)保存 2、6、12、2 4、72h。保存后将每组中的 5个同种带瓣大动脉浸入 30 0 μl含重组腺病毒 (Ad5CMVLacZ)的DMEM液中 ,而另1个则浸入 30 0 μl不含重组腺病毒的DMEM液中 ,均在室温下放置 30min ,然后分别移植于 15只成兔受体的两侧颈动脉。术后 5d取出移植物 ,进行组织化学染色。结果 所有转染重组腺病毒的同种带瓣大动脉均有不同程度的基因表达 ,Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组阳性细胞平均表达量分别 42 .6±8.5、38.2± 12 .7、34 .5± 10 .2、31.6± 9.6、36.4± 13.2 ;组间转基因细胞表达量差异无显著性 (P >0 .0 5 )。对照组均为阴性。结论 对低温保存不同时间后的同种带瓣大动脉进行转基因是可行的。

Objective To assess the feasibility of g ene transfer to homograft conduits after cold preservation.Methods Thirty homograft conduits of infant rabbi ts were divided into five groups and then stored in M199 for cold preservation a t 4℃ for 2, 6, 12, 24 and 72 h respectively. Five grafts of each group were in cubated in 300μl DMEM containing adenovirus carrying theβ-galactosidas gene , and one of each group was incubated in 300μl DMEM not containing adenovirus . After 30min incubation, bilateral aorto-cartid transplantation was performe d in 15 recipient adult rabbits. The homografts were harvested at 5 days after t ransplantation and assayed for β-galactosidase activity. Results Successful gene transfer and expression of the β-gal gene were detected in all homograft conduits. The mean total numbers of positively stained cells for β-galactosidas activity were 42.6±8.5 in the gr oup Ⅰ, 38.2±12.7 in the group Ⅱ,34.5±10.2 in the group Ⅲ, 31.6±9.6 in the group Ⅳ and 36.4±13.2 in the group Ⅴ respectively. No significant differences among the groups were found (P>0.05). No transduction was found in the control group. Conclusion It is feasible to perform adenovira l-mediated gene transfer to homograft conduits after cold preservation.