title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro.
abstract: The purpose of this study was to establish a method for monitoring the neural differentiation of stem cells using ferritin transgene expression, under the control of a neural-differentiation-inducible promoter, and magnetic resonance imaging (MRI). Human adipose tissue-derived mesenchymal stem cells (hADMSCs) were transduced with a lentivirus containing the human ferritin heavy chain 1 (FTH1) gene coupled to one of three neural cell-specific promoters: human synapsin 1 promoter (SYN1p, for neurons), human glial fibrillary acidic protein promoter (GFAPp, for astrocytes), and human myelin basic protein promoter (MBPp, for oligodendrocytes). Three groups of neural-differentiation-inducible ferritin-expressing (NDIFE) hADMSCs were established: SYN1p-FTH1, GFAPp-FTH1, and MBPp-FTH1. The proliferation rate of the NDIFE hADMSCs was evaluated using a Cell Counting Kit-8 assay. Ferritin expression was assessed with western blotting and immunofluorescent staining before and after the induction of differentiation in NDIFE hADMSCs. The intracellular iron content was measured with Prussian blue iron staining and inductively coupled plasma mass spectrometry. R2 relaxation rates were measured with MRI in vitro. The proliferation rates of control and NDIFE hADMSCs did not differ significantly (P > 0.05). SYN1p-FTH1, GFAPp-FTH1, and MBPp-FTH1 hADMSCs expressed specific markers of neurons, astrocytes, and oligodendrocytes, respectively, after neural differentiation. Neural differentiation increased ferritin expression twofold, the intracellular iron content threefold, and the R2 relaxation rate two- to threefold in NDIFE hADMSCs, resulting in notable hypointensity in T2-weighted images (P < 0.05). These results were cross-validated. Thus, a link between neural differentiation and MRI signals (R2 relaxation rate) was established in hADMSCs. The use of MRI and neural-differentiation-inducible ferritin expression is a viable method for monitoring the neural differentiation of hADMSCs.
authors.affiliation: Department of Anatomy, Institute of Clinical Anatomy, Southern Medical University, Guangzhou, China.
authors.forename: Chengang
authors.initials: C
authors.lastname: Song
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, China; Department of Pathology, Shenzhen Affiliated Hospital, Guangzhou University of Traditional Chinese Medicine, Shenzhen, China.
authors.forename: Jiachuan
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, China.
authors.forename: Cuiping
authors.initials: C
authors.lastname: Mo
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, China.
authors.forename: Shuhua
authors.initials: S
authors.lastname: Mu
authors.affiliation: Department of Pathology, Southern Medical University, Guangzhou, China.
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou, China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Anatomy, Institute of Clinical Anatomy, Southern Medical University, Guangzhou, China.
authors.forename: Shizhen
authors.initials: S
authors.lastname: Zhong
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, China.
authors.forename: Zhenfu
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, China.
authors.forename: Guangqian
authors.initials: G
authors.lastname: Zhou
mesh_terms: D002478:Cells, Cultured; D005293:Ferritins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008279:Magnetic Resonance Imaging; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D055495:Neurogenesis; D011401:Promoter Regions, Genetic; D015533:Transcriptional Activation
chemical_list: D005293:Ferritins