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在梗死心脏中利用可注射支架对脂肪组织来源干细胞进行实时追踪。

Real-time tracking of adipose tissue-derived stem cells with injectable scaffolds in the infarcted heart.

作者信息

Yang Jun-jie, Liu Zhi-qiang, Zhang Jin-ming, Wang Hai-bin, Hu Shun-yin, Liu Jian-feng, Wang Chang-yong, Chen Yun-dai

机构信息

Department of Cardiology, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing, 100853, People's Republic of China.

出版信息

Heart Vessels. 2013 May;28(3):385-96. doi: 10.1007/s00380-012-0275-0. Epub 2012 Sep 1.

Abstract

Adipose tissue-derived stem cells (ADSCs) has shown promise in the emerging field of regenerative medicine. Many studies have highlighted the importance of coadministering a "scaffold" for increasing intramyocardial retention of stem cells. In this work, an optimized method was developed for efficient transduction of ADSCs with a lentiviral vector carrying a triple-fusion reporter gene that consists of firefly luciferase, monomeric red fluorescence protein, and truncated thymidine kinase (fluc-mrfp-ttk). The transduced ADSCs were assessed on biological performance and transplanted into infarcted heart with fibrin scaffolds. In vivo cell retention was tracked by bioluminescence imaging (BLI) and micro positron emission tomography/computed tomography (PET/CT) imaging. Histological assessment was performed for regeneration potentials. The results showed that lentiviral transduction did not influence cell functions. In vitro imaging analysis showed a robust linear correlation between cell numbers and BLI signals (R (2) = 0.99) as well as between cell numbers and radiotracer uptakes (R (2) = 0.98). Transduced ADSCs were visualized in the heart under both BLI and PET/CT imaging, contributing to cardiomyocyte regeneration and angiogenesis in the implanted areas. Compared with BLI monitoring, PET/CT data provided precise localization for cell retention. Thus, a combination of imaging modalities can assist in reliable and efficient monitoring of transplanted cells, holding great potential for the transplantation of injectable scaffolds encapsulating stem cells in treating heart disease.

摘要

脂肪组织来源的干细胞(ADSCs)在新兴的再生医学领域已展现出前景。许多研究强调了共同给予“支架”以增加干细胞心肌内滞留的重要性。在这项工作中,开发了一种优化方法,用于用携带由萤火虫荧光素酶、单体红色荧光蛋白和截短的胸苷激酶组成的三融合报告基因(fluc-mrfp-ttk)的慢病毒载体高效转导ADSCs。对转导后的ADSCs进行生物学性能评估,并将其与纤维蛋白支架一起移植到梗死心脏中。通过生物发光成像(BLI)和微型正电子发射断层扫描/计算机断层扫描(PET/CT)成像追踪体内细胞滞留情况。进行组织学评估以确定再生潜力。结果表明,慢病毒转导不影响细胞功能。体外成像分析显示细胞数量与BLI信号之间(R(2)= 0.99)以及细胞数量与放射性示踪剂摄取之间(R(2)= 0.98)存在很强的线性相关性。在BLI和PET/CT成像下均可在心脏中观察到转导后的ADSCs,它们有助于植入区域的心肌细胞再生和血管生成。与BLI监测相比,PET/CT数据为细胞滞留提供了精确的定位。因此,成像方式的组合可有助于可靠且高效地监测移植细胞,在治疗心脏病时将封装干细胞的可注射支架进行移植具有巨大潜力。

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