Assessment of biological characteristics of adipose tissue‑derived stem cells co‑labeled with Molday ION Rhodamine B™ and green fluorescent protein in vitro

Nan,Hua; Huang,Jiacheng; Li,Hongmian; Li,Qiong; Liu,Dalie

doi:10.3892/mmr.2013.1694

Assessment of biological characteristics of adipose tissue‑derived stem cells co‑labeled with Molday ION Rhodamine B™ and green fluorescent protein in vitro

Authors:
- Hua Nan
- Jiacheng Huang
- Hongmian Li
- Qiong Li
- Dalie Liu
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Affiliations: Plastic Surgery Institute, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China, Plastic Surgery Institute, Zhongshan Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510275, P.R. China
Published online on: September 18, 2013 https://doi.org/10.3892/mmr.2013.1694
Pages: 1446-1452

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Abstract

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.

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1	De Ugarte DA, Morizono K, Elbarbary A, et al: Comparison of multi-lineage cells from human adipose tissue and bone marrow. Cells Tissues Organs. 174:101–109. 2003.PubMed/NCBI
2	Bauer G, Dao MA, Case SS, et al: In vivo biosafety model to assess the risk of adverse events from retroviral and lentiviral vectors. Mol Ther. 16:1308–1315. 2008. View Article : Google Scholar : PubMed/NCBI
3	Bulte JW and Kraitchman DL: Iron oxide MR contrast agents for molecular and cellular imaging. NMR Biomed. 17:484–499. 2004. View Article : Google Scholar : PubMed/NCBI
4	Addicott B, Willman M, Rodriguez J, Padgett K, Han D, Berman D, Hare JM and Kenyon NS: Mesenchymal stem cell labeling and in vitro MR characterization at 1.5T of new SPIO contrast agent: Molday ION Rhodamine-B™. Contrast Media Mol Imaging. 6:7–18. 2011.PubMed/NCBI
5	Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP and Hedrick MH: Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 7:211–228. 2001. View Article : Google Scholar : PubMed/NCBI
6	Caplan AI: Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics (Review). Tissue Eng. 11:1198–1211. 2005. View Article : Google Scholar : PubMed/NCBI
7	Dyson SC and Barker RA: Cell-based therapies for Parkinson’s disease. Expert Rev Neurother. 11:831–844. 2011.
8	Ward TH and Lippincott-Schwartz J: The uses of green fluorescent protein in mammalian cells. Methods Biochem Anal. 47:305–337. 2006.PubMed/NCBI
9	Serganova I, Mayer-Kukuck P, Huang R and Blasberg R: Molecular imaging: reporter gene imaging. Handb Exp Pharmacol. 185:167–223. 2008. View Article : Google Scholar
10	Zhu XM, Wang YX, Leung KC, Lee SF, Zhao F, Wang DW, Lai JM, Wan C, Cheng CH and Ahuja AT: Enhanced cellular uptake of aminosilane-coated superparamagnetic iron oxide nanoparticles in mammalian cell lines. Int J Nanomedicine. 7:953–964. 2012.PubMed/NCBI
11	Lee ES, Chan J, Shuter B, et al: Microgel iron oxide nanoparticles for tracking human fetal mesenchymal stem cells through magnetic resonance imaging. Stem Cells. 27:1921–1931. 2009. View Article : Google Scholar : PubMed/NCBI
12	Domen J and Weissman IL: Self-renewal, differentiation or death: regulation and manipulation of hematopoietic stem cell fate. Mol Med Today. 5:201–208. 1999. View Article : Google Scholar : PubMed/NCBI
13	Kraitchman DL, Heldman AW, Atalar E, Amado LC, Martin BJ, Pittenger MF, Hare JM and Bulte JW: In vivo magnetic resonance imaging of mesenchymal stem cells in myocardial infarction. Circulation. 107:2290–2293. 2003. View Article : Google Scholar : PubMed/NCBI
14	Zhu Y, Liu T, Song K, Fan X, Ma X and Cui Z: Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochem Funct. 26:664–675. 2008. View Article : Google Scholar : PubMed/NCBI
15	Taléns-Visconti R, Bonora A, Jover R, Mirabet V, Carbonell F, Castell JV and Gómez-Lechón MJ: Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells. World J Gastroenterol. 12:5834–5845. 2006.PubMed/NCBI
16	Wang L, Deng J, Wang J, et al: Superparamagnetic iron oxide does not affect the viability and function of adipose-derived stem cells, and superparamagnetic iron oxide-enhanced magnetic resonance imaging identifies viable cells. Magn Reson Imaging. 27:108–119. 2009. View Article : Google Scholar : PubMed/NCBI
17	Ren Z, Wang J, Zou C, Guan Y and Zhang YA: Labeling of cynomolgus monkey bone marrow-derived mesenchymal stem cells for cell tracking by multimodality imaging. Sci China Life Sci. 54:981–987. 2011. View Article : Google Scholar : PubMed/NCBI
18	Arbab AS, Yocum GT, Rad AM, Khakoo AY, Fellowes V, Read EJ and Frank JA: Labeling of cells with ferumoxides-protamine sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells. NMR Biomed. 18:553–559. 2005. View Article : Google Scholar : PubMed/NCBI
19	Acton PD and Zhou R: Imaging reporter genes for cell tracking with PET and SPECT. Q J Nucl Med Mol Imaging. 49:349–360. 2005.PubMed/NCBI
20	Higuchi T, Anton M, Dumler K, et al: Combined reporter gene PET and iron oxide MRI for monitoring survival and localization of transplanted cells in the rat heart. J Nucl Med. 50:1088–1094. 2009. View Article : Google Scholar : PubMed/NCBI
21	Qiu B, Treuting P, Zhan X, Xie D, Frevert CW and Yang X: Dual transfer of GFP gene and MGd into stem-progenitor cells: toward in vivo MRI of stem cell-mediated gene therapy of atherosclerosis. Acad Radiol. 17:547–552. 2010. View Article : Google Scholar : PubMed/NCBI