MRI detection of the malignant transformation of stem cells through reporter gene expression driven by a tumor-specific promoter

Abstract Background Existing evidence has shown that mesenchymal stem cells (MSCs) can undergo malignant transformation, which is a serious limitation of MSC-based therapies. Therefore, it is necessary to monitor malignant transformation of MSCs via a noninvasive imaging method. Although reporter gene-based magnetic resonance imaging (MRI) has been successfully applied to longitudinally monitor MSCs, this technique cannot distinguish the cells before and after malignant transformation. Herein, we investigated the feasibility of using a tumor-specific promoter to drive reporter gene expression for MRI detection of the malignant transformation of MSCs. Methods The reporter gene ferritin heavy chain (FTH1) was modified by adding a promoter from the tumor-specific gene progression elevated gene-3 (PEG3) and transduced into MSCs to obtain MSCs-PEG3-FTH1. Cells were induced to undergo malignant transformation via indirect coculture with C6 glioma cells, and these transformed cells were named MTMSCs-PEG3-FTH1. Western blot analysis of FTH1 expression, Prussian blue staining and transmission electron microscopy (TEM) to detect intracellular iron, and MRI to detect signal changes were performed before and after malignant transformation. Then, the cells before and after malignant transformation were inoculated subcutaneously into nude mice, and MRI was performed to observe the signal changes in the xenografts. Results After induction of malignant transformation, MTMSCs demonstrated tumor-like features in morphology, proliferation, migration, and invasion. FTH1 expression was significantly increased in MTMSCs-PEG3-FTH1 compared with MSCs-PEG3-FTH1. Prussian blue staining and TEM showed a large amount of iron particles in MTMSCs-PEG3-FTH1 but a minimal amount in MSCs-PEG3-FTH1. MRI demonstrated that the T2 value was significantly decreased in MTMSCs-PEG3-FTH1 compared with MSCs-PEG3-FTH1. In vivo, mass formation was observed in the MTMSCs-PEG3-FTH1 group but not the MSCs-PEG3-FTH1 group. T2-weighted MRI showed a significant signal decrease, which was correlated with iron accumulation in the tissue mass. Conclusions We developed a novel MRI model based on FTH1 reporter gene expression driven by the tumor-specific PEG3 promoter. This approach could be applied to sensitively detect the occurrence of MSC malignant transformation.

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Hemato-endothelial differentiation from lentiviral-transduced human embryonic stem cells retains durable reporter gene expression under the control of ubiquitin promoter

Cytotechnology ◽

10.1007/s10616-010-9258-0 ◽

2010 ◽

Vol 62 (1) ◽

pp. 31-42 ◽

Cited By ~ 5

Author(s):

Hua Jiang ◽

Xiaolong Lin ◽

Youji Feng ◽

Yi Xie ◽

Jinlan Han ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Embryonic Stem Cells ◽

Reporter Gene ◽

Human Embryonic Stem Cells ◽

Embryonic Stem ◽

Reporter Gene Expression ◽

Endothelial Differentiation ◽

Ubiquitin Promoter

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oct4 -EGFP reporter gene expression marks the stem cells in embryonic development and in adult gonads of transgenic medaka

Molecular Reproduction and Development ◽

10.1002/mrd.22135 ◽

2012 ◽

Vol 80 (1) ◽

pp. 48-58 ◽

Cited By ~ 17

Author(s):

Alexander Froschauer ◽

Mst. Muslima Khatun ◽

David Sprott ◽

Alexander Franz ◽

Christiane Rieger ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Embryonic Development ◽

Reporter Gene ◽

Reporter Gene Expression ◽

Egfp Reporter Gene ◽

Egfp Reporter

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In VitroNeural Differentiation of Bone Marrow Mesenchymal Stem Cells Carrying the FTH1 Reporter Gene and Detection with MRI

BioMed Research International ◽

10.1155/2018/1978602 ◽

2018 ◽

Vol 2018 ◽

pp. 1-14 ◽

Cited By ~ 3

Author(s):

Tong Mu ◽

Yong Qin ◽

Bo Liu ◽

Xiaoya He ◽

Yifan Liao ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Differentiation ◽

Reporter Gene ◽

Neural Differentiation ◽

Neuron Specific Enolase ◽

Stem Cell Differentiation ◽

Blue Staining ◽

Differentiated Cells

Magnetic resonance imaging (MRI) based on the ferritin heavy chain 1 (FTH1) reporter gene has been used to trace stem cells. However, whether FTH1 expression is affected by stem cell differentiation or whether cell differentiation is affected by reporter gene expression remains unclear. Here, we explore the relationship between FTH1 expression and neural differentiation in the differentiation of mesenchymal stem cells (MSCs) carrying FTH1 into neuron-like cells and investigate the feasibility of using FTH1 as an MRI reporter gene to detect neurally differentiated cells. By inducing cell differentiation with all-trans retinoic acid and a modified neuronal medium, MSCs and MSCs-FTH1 were successfully differentiated into neuron-like cells (Neurons and Neurons-FTH1), and the neural differentiation rates were (91.56±7.89)% and (92.23±7.64)%, respectively. Neuron-specific markers, including nestin, neuron-specific enolase, and microtubule-associated protein-2, were significantly expressed in Neurons-FTH1 and Neurons without noticeable differences. On the other hand, FTH1 was significantly expressed in MSCs-FTH1 and Neurons-FTH1 cells, and the expression levels were not significantly different. The R2 value was significantly increased in MSCs-FTH1 and Neurons-FTH1 cells, which was consistent with the findings of Prussian blue staining, transmission electron microscopy, and intracellular iron measurements. These results suggest that FTH1 gene expression did not affect MSC differentiation into neurons and was not affected by neural differentiation. Thus, MRI reporter gene imaging based on FTH1 can be used for the detection of neurally differentiated cells from MSCs.

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Serial Noninvasive In Vivo Positron Emission Tomographic Tracking of Percutaneously Intramyocardially Injected Autologous Porcine Mesenchymal Stem Cells Modified for Transgene Reporter Gene Expression

Circulation Cardiovascular Imaging ◽

10.1161/circimaging.108.797449 ◽

2008 ◽

Vol 1 (2) ◽

pp. 94-103 ◽

Cited By ~ 105

Author(s):

Mariann Gyöngyösi ◽

Jeronimo Blanco ◽

Teréz Marian ◽

Lajos Trón ◽

Örs Petneházy ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Reporter Gene ◽

Reporter Gene Expression ◽

Positron Emission Tomographic ◽

Positron Emission

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SHED Cells Labeled with MIRB can be Tracked in Repairing Periodontal Bone Defects in Rats

10.21203/rs.3.rs-111231/v1 ◽

2020 ◽

Author(s):

Nan Zhang ◽

Li Xu ◽

Hao Song ◽

Chunqing Bu ◽

Jie Kang ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Prussian Blue ◽

Bone Defect ◽

Immunohistochemical Analysis ◽

Blue Staining ◽

Iron Oxide Particles ◽

Defect Model ◽

Prussian Blue Staining

Abstract Background: Chronic periodontitis could lead to alveolar bone resorption and even tooth loss. Stem cells from exfoliated deciduous teeth (SHED) are the proper seed cells for bone regeneration because of their potential in osteogenic differentiation. However tracking the survival, migration and differentiation of the transplanted stem cells is necessary to improve the transplantation success. Methods: Superparamagnetic iron oxide particles (SPIO) Molday ION Rhodamine-BTM (MIRB) were used for labeling and monitoring SHED cells in vivo by magnetic resonance imaging (MRI). Proper labeling concentration of MIRB was determined by cell viability, proliferation, osteogenic differentiation and MRI analysis in vitro after SHED cells were labeled with MIRB at different concentration of 12.5, 25, 50, 100μgFe/mL. MIRB labeled SHED were transplanted to the periodontal bone defect model in rats and tracked by MRI in vivo. The regeneration of periodontal bone were calculated with HE and immunohistochemical analysis. The survival of transplanted SHED cells in vivo was verified with Prussian blue staining.Results: After testing 25μg Fe/mL MIRB was used in vivo cells tracking. After transplanted to the periodontal bone defect model in rats, the MIRB labeled SHED could be tracked in vivo through the artifact of the low intensity signal caused by Fe3+ at 6 and 9 weeks post-surgery. HE and immunohistochemical analysis showed that both SHED labeled and unlabeled with MIRB could promote regeneration of periodontal bone defect. Prussian blue staining further verified the survival of transplanted SHED cells in vivo. Conclusions: Overall, SHED cells could promote the regeneration of periodontal bone in rats and the survival of SHED cells could be tracked by labeling with MIRB in vivo. However the distribution of the positive cells at the edge of the regenerated new bone remind us the SHED cell could promote the regeneration of new bone by factors section.

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