Plastic Surgery Institute, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China.
Mol Med Rep. 2013 Nov;8(5):1446-52. doi: 10.3892/mmr.2013.1694. Epub 2013 Sep 18.
The current study aimed to investigate adipose tissue-derived stem cells (ADSCs) in vivo by multimodality imaging following implantation for cellular therapy. The biological characteristics of ADSCs co-labeled with Molday ION Rhodamine B™ (MIRB) and green fluorescent protein (GFP) were studied in vitro. Following rat ADSC isolation and culture, a combined labeling strategy for ADSCs based on genetic modification of the reporter gene GFP with lentiviral vector expression enhancement and physical MIRB labeling was performed. Cell viability, proliferation, membrane-bound antigens and multiple differentiation ability were compared between the labeled and unlabeled ADSCs. The ADSCs were successfully labeled with GFP and MIRB, showing various fluorescent colors for marker identification. The fluorescence emitted by the GFP protein was sustained and exhibited stable expression, while MIRB fluorescence decreased with time. Compared with the unlabeled ADSCs, no significant differences were detected in cell viability, proliferation, membrane-bound antigens and multiple differentiation ability in the co-labeled samples (P>0.05). No significant effects on the biophysical properties of ADSCs were observed following co-labeling with lentiviral vectors encoding the gene for emerald green fluorescent protein and MIRB. The ADSCs were able to be efficiently tracked in vitro and in vivo by multimodality imaging thus, the co-labeling approach provides a novel strategy for therapeutic gene studies.
本研究旨在通过植入细胞治疗后的多模态成像来研究脂肪组织来源的干细胞(ADSCs)。在体外研究了共标记有 Molday ION Rhodamine BTM(MIRB)和绿色荧光蛋白(GFP)的 ADSCs 的生物学特性。在大鼠 ADSC 分离和培养后,采用基于慢病毒载体表达增强和物理 MIRB 标记的报告基因 GFP 遗传修饰的联合标记策略对 ADSC 进行了标记。比较了标记和未标记的 ADSC 之间的细胞活力、增殖、膜结合抗原和多向分化能力。ADSCs 成功地被 GFP 和 MIRB 标记,显示出不同的荧光颜色用于标记识别。GFP 蛋白发出的荧光持续存在,表现出稳定的表达,而 MIRB 荧光随时间降低。与未标记的 ADSC 相比,共标记样品中的细胞活力、增殖、膜结合抗原和多向分化能力没有显著差异(P>0.05)。共标记编码 Emerald 绿色荧光蛋白和 MIRB 的基因的慢病毒载体对 ADSC 的生物物理特性没有显著影响。通过多模态成像,ADSCs 能够在体外和体内被高效追踪,因此,共标记方法为治疗基因研究提供了一种新策略。