摘要
骨髓机制细胞(BMSCs)已经被证明对多种人类疾病和损伤治疗有用。然而,他们的修复能力限制于他们微弱的迁移和归巢能力,主要依赖于SDF-1/CXCR4 轴。大多数BMSCs传代培养缺乏细胞表面CXCR4受体表达及表现出受损的迁移能力。为了增加SDF-1响应能力和促进细胞迁移及培养的BMSCs的存活,我们采用超声靶向微泡破坏(UTMD)与脂质体结合来增加CXCR4体内表达。我们分离和培养了第三代大鼠BMSCs,采用微泡造影剂介导的超声辐射和脂质体,转导他们到重组质粒pDsRed-CXCR4。与一些病毒载体相比,我们在此陈述的方法有较好的转染效率,CXCR4表达及技术重现性。这种方法的益处可能是由于震荡波引起的“声孔效应”与由UTMD 产生的空穴现象导致的微射流的结合。转染之后,我们进行了转板迁移测定分析,发现CXCR4介导的BMSCs 的转移能力比对照组高出9倍。我们在此描述的方法为CXCR4的高水平表达提供了一种简单安全非病毒载体方式。这与增强的传代培养BMSCs的迁移有关,也许对临床应用有用。
关键词: 骨髓机制细胞,细胞迁移,CXCR4,脂质体,微细气泡,超声波
Current Gene Therapy
Title:Transfection of CXCR-4 Using Microbubble-Mediated Ultrasound Irradiation and Liposomes Improves the Migratory Ability of Bone Marrow Stromal Cells
Volume: 15 Issue: 1
Author(s): Gong Wang, Zhongxiong Zhuo, Qian Zhang, Yali Xu, Shengzheng Wu, Lu Li, Hongmei Xia and Yunhua Gao
Affiliation:
关键词: 骨髓机制细胞,细胞迁移,CXCR4,脂质体,微细气泡,超声波
摘要: Bone marrow stromal cells (BMSCs) have proven useful for the treatment of various human diseases and injuries. However, their reparative capacity is limited by their poor migration and homing ability, which are primarily dependent on the SDF-1/CXCR4 axis. Most subcultured BMSCs lack CXCR4 receptor expression on the cell surface and exhibit impaired migratory capacity. To increase responsiveness to SDF-1 and promote cell migration and survival of cultured BMSCs, we used a combination of ultrasound-targeted microbubble destruction (UTMD) and liposomes to increase CXCR4 expression in vitro. We isolated and cultured rat BMSCs to their third passage and transduced them with recombinant plasmid pDsRed-CXCR4 using microbubble-mediated ultrasound irradiation and liposomes. Compared to some viral vectors, the method we employed here resulted in significantly better transfection efficiency, CXCR4 expression, and technical reproducibility. The benefits of this approach are likely due to the combination of “sonoporation” caused by shockwaves and microjet flow resulting from UTMD-generated cavitation. Following transfection, we performed a transwell migration assay and found that the migration ability of CXCR4-modified BMSCs was 9-fold higher than controls. The methods we describe here provide an effective, safe, non-viral means to achieve high levels of CXCR4 expression. This is associated with enhanced migration of subcultured BMSCs and may be useful for clinical application as well.
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Gong Wang, Zhongxiong Zhuo, Qian Zhang, Yali Xu, Shengzheng Wu, Lu Li, Hongmei Xia and Yunhua Gao , Transfection of CXCR-4 Using Microbubble-Mediated Ultrasound Irradiation and Liposomes Improves the Migratory Ability of Bone Marrow Stromal Cells, Current Gene Therapy 2015; 15 (1) . https://dx.doi.org/10.2174/1566523214666141121111220
DOI https://dx.doi.org/10.2174/1566523214666141121111220 |
Print ISSN 1566-5232 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5631 |
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