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. 2015 Aug 6;10(8):e0135111.
doi: 10.1371/journal.pone.0135111. eCollection 2015.

Neuronal Differentiation of Human Mesenchymal Stem Cells Using Exosomes Derived from Differentiating Neuronal Cells

Yuji S Takeda  1 , Qiaobing Xu  1
Affiliations

Neuronal Differentiation of Human Mesenchymal Stem Cells Using Exosomes Derived from Differentiating Neuronal Cells

Yuji S Takeda et al. PLoS One. .

Abstract

Exosomes deliver functional proteins and genetic materials to neighboring cells, and have potential applications for tissue regeneration. One possible mechanism of exosome-promoted tissue regeneration is through the delivery of microRNA (miRNA). In this study, we hypothesized that exosomes derived from neuronal progenitor cells contain miRNAs that promote neuronal differentiation. We treated mesenchymal stem cells (MSCs) daily with exosomes derived from PC12 cells, a neuronal cell line, for 1 week. After the treatment with PC12-derived exosomes, MSCs developed neuron-like morphology, and gene and protein expressions of neuronal markers were upregulated. Microarray analysis showed that the expression of miR-125b, which is known to play a role in neuronal differentiation of stem cells, was much higher in PC12-derived exosomes than in exosomes from B16-F10 melanoma cells. These results suggest that the delivery of miRNAs contained in PC12-derived exosomes is a possible mechanism explaining the neuronal differentiation of MSC.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Schematic representation of this study.
Exosomes were collected from the conditioned media of PC12 cells differentiating into neuron-like cells. Human mesenchymal stem cells (hMSC) were dosed with the exosomes, and then evaluated for differentiation.
Fig 2
Fig 2. Size and morphology of exosomes.
(A) Typical size distribution for an exosome sample derived from PC12 cells, measured with dynamic light scattering (DLS). (B) DLS analysis of exosome samples. n = 9. All error bars represent standard deviation. (C) Transmission electron microscopy (TEM) image of a typical exosome sample derived from B16-F10 cells. Scale bar, 100 nm.
Fig 3
Fig 3. Cell morphology of hMSC after treatment with exosomes from neuronal cells.
hMSCs were treated for 1 week with exosomes derived from differentiating PC12 cells. MAP2, NSE, and 160 kDa neurofilament (NF160) were immunostained. As negative controls, non-treated hMSCs and hMSCs treated with exosomes derived from B16-F10 cells are shown. Blue: cell nuclei. Scale bar, 50 μm.
Fig 4
Fig 4. mRNA expression of hMSC treated with exosome samples for 1 week.
(A) MAP2, (B) NSE. **: p < 0.01, ***: p < 0.001. Ratio compared with non-treated sample. All error bars represent standard deviation.
Fig 5
Fig 5. Western blot of neuronal marker proteins (NSE, MAP2, and GAPDH).
(A) MSCs were treated with exosomes for 1 week. (B) PC12 exosomes and B16-F10 cells treated with PC12 exosomes. Cell lysate from differentiated PC12 cells, treated with NGF for 1 week, was used as a positive control.
Fig 6
Fig 6. miRNA expression in exosomes.
(A) 101 total probes common between rat and mouse species and detectable in at least 1 sample were used in analysis by hierarchical clustering. Intensities for the probes were log-2-transformed. (B, C) Comparison of miRNA expression between D9 and D0 exosome (B), and D9 and B16-F10 exosome (C).
See this image and copyright information in PMC

References

    1. Straley KS, Foo CW, Heilshorn SC. Biomaterial design strategies for the treatment of spinal cord injuries. J Neurotrauma. 2010;27: 1–19. 10.1089/neu.2009.0948 - DOI - PMC - PubMed
    1. Tabesh H, Amoabediny G, Nik NS, Heydari M, Yosefifard M, Siadat SO, et al. The role of biodegradable engineered scaffolds seeded with Schwann cells for spinal cord regeneration. Neurochem Int. 2009;54: 73–83. 10.1016/j.neuint.2008.11.002 - DOI - PubMed
    1. Willerth SM, Sakiyama-Elbert SE. Approaches to neural tissue engineering using scaffolds for drug delivery. Adv Drug Deliv Rev. 2007;59: 325–338. 10.1016/j.addr.2007.03.014 - DOI - PMC - PubMed
    1. Kumar AA, Kumar SR, Narayanan R, Arul K, Baskaran M. Autologous bone marrow derived mononuclear cell therapy for spinal cord injury: A phase I/II clinical safety and primary efficacy data. Exp Clin Transplant. 2009;7: 241–248. - PubMed
    1. Sharma A, Gokulchandran N, Chopra G, Kulkarni P, Lohia M, Badhe P, et al. Administration of autologous bone marrow-derived mononuclear cells in children with incurable neurological disorders and injury is safe and improves their quality of life. Cell Transplant. 2012;21 Suppl 1: S79–90. 10.3727/096368912X633798 - DOI - PubMed

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