Blood-derived smooth muscle cells as a target for gene delivery.
Blood-derived smooth muscle cells as a target for gene delivery.
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血液来源的平滑肌细胞作为基因传递的靶标。
DOI：
10.1016/j.jvs.2007.10.039
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发表时间：
2008
影响因子：
4.3
通讯作者：
Yu,Hong
中科院分区：
文献类型：
--
作者：
Yang,Zhe;Shao,Hongwei;Tan,Yaohong;Eton,Darwin;Yu,Hong
目的探讨利用血源性平滑肌细胞(BD-SMCs)作为靶点输送治疗性蛋白的可行性。在含有血小板衍生生长因子BB的培养液中，将MNC培养的生长集落分化为BD-SMC。免疫细胞化学检测BD-SMC的表型特征。以血管来源的SMC(VSMCs)为对照，对BD-SMC的细胞增殖、逆转录病毒载体的基因转移效率、细胞凋亡率和转导基因产物的生物学活性进行了体内外检测。BD-SMCs和VSMCs在细胞增殖、迁移、黏附和基因转移效率方面无显著差异。用携带分泌型碱性磷酸酶基因的逆转录病毒载体转导BD-SMC后，24小时内每106细胞产生174±50μg生物活性的分泌型碱性磷酸酶基因。静脉注射5×106细胞后，3天后循环中SEAP浓度由0.14±0.0 4μg/ml上升至2.34±0.16μg/ml(P<0.0 1)。1周后循环中SEAP降至1.76μg/ml，8周后一直维持在该水平，12周时降至细胞注射前水平。结论VSMC与成熟VSMC具有相似的生物学特性，可作为基因转移的新靶点。
OBJECTIVETo examine the feasibility of using blood-derived smooth muscle cells (BD-SMCs) as a target for to deliver therapeutic proteins.MATERIALS AND METHODSMononuclear cells (MNC) were isolated from peripheral blood. The outgrowth colonies from MNC culture were differentiated into BD-SMCs in media containing platelet-derived growth factor BB. Phenotypic characterization of BD-SMCs was assessed by immunocytochemistry. Cell proliferation, gene transfer efficiency with a retroviral vector, apoptosis, and the biological activity of the transduced gene product from the BD-SMCs were evaluated in vitro and in vivo in comparison with vascular derived SMC (VSMCs).RESULTSBD-SMCs stained positive for SMC markers. No significant difference was observed between BD-SMCs and VSMCs in cell proliferation, migration, adhesiveness, and gene transfer efficiency. After BD-SMCs were transduced with a retroviral vector carrying the secreted alkaline phosphatase gene (SEAP), 174 ± 50 μg biologically active SEAP was produced per 106cells over 24 hours. After injecting 5 × 106cells expressing SEAP intravenously into rabbits, SEAP concentration increased significantly in the circulation from 0.14 ± 0.04 μg/ml to 2.34 ± 0.16 μg/ml 3 days after cell injection (P < .01, n = 3). Circulating levels of SEAP decreased to 1.76 μg /ml 1 week later and remained at this level up to 8 weeks, then declined to pre-cell injection level at 12 weeks. VSMC in vivo gene expression data were equivalent.CONCLUSIONBD-SMCs have similar characteristics to mature VSMCs and can be used as a novel target for gene transfer to deliver a therapeutic protein.