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目的 探讨腺病毒介导的人骨形态发生蛋白 (BMP 2 )转染对体外培养骨髓间质干细胞(MSCs)成骨能力的影响。 方法 取日本大耳白兔 2 0只自双侧股骨大转子取材培养MSCs ,左侧来源细胞为实验组 ,右侧为对照组。以复制缺陷重组腺病毒介导人BMP 2基因转染MSCs后 ,用ALP检测两组细胞的ALP活性 ;骨钙素RT PCR检测两组细胞Ⅰ型胶原的表达 ;BGP放免分析检测两组细胞的骨钙素含量。 结果 转基因组和对照组ALP分泌量 (U/L)分别为 7 0 1± 0 5 9、5 2 3± 0 5 5 ;RT PCRⅠ型胶原的表达为 1 3 5± 0 12、3 65± 0 3 7;骨钙素 (ng/ml)为 2 4 5 0± 0 93和 15 45± 1 81。两组间差异均有显著性 (P <0 0 5 )。 结论 用腺病毒介导人BMP 2蛋白作用后的MSCs具有成骨细胞的生物学特性。腺病毒介导人BMP 2转基因可以提高MSCs的体外成骨能力。
Objective To investigate the effect of adenovirus-mediated transfection of human bone morphogenetic protein (BMP 2) on the osteogenic ability of cultured bone marrow mesenchymal stem cells (MSCs) in vitro. Methods Twenty Japanese white rabbits were harvested from bilateral femoral trochanter. MSCs were cultured on the left and the control group on the right. ALP was used to detect ALP activity in MSCs transfected with recombinant adenovirus-mediated human BMP-2 gene; osteocalcin RT-PCR was used to detect the expression of type I collagen in the two groups of cells; Osteocalcin content. Results The ALP secretion (U / L) of the transgenic group and the control group was 701 ± 0 5 9 and 52 3 ± 0 5 5, respectively. The expression of RT-PCR typeⅠcollagen was 1 35 ± 0 12,3 65 ± 0 3 7; osteocalcin (ng / ml) was 2450 ± 0 93 and 15 45 ± 1 81. The differences between the two groups were significant (P <0 05). CONCLUSION: MSCs cultured with adenovirus-mediated human BMP-2 have the biological characteristics of osteoblasts. Adenovirus-mediated human BMP2 transgene can enhance MSCs in vitro osteogenic potential.