PDGF, NT-3 and IGF-2 in Combination Induced Transdifferentiation of Muscle-Derived Stem Cells into Schwann Cell-Like Cells

PLOS ONE, Dec 2019

Muscle-derived stem cells (MDSCs) are multipotent stem cells with a remarkable long-term self-renewal and regeneration capacity. Here, we show that postnatal MDSCs could be transdifferentiated into Schwann cell-like cells upon the combined treatment of three neurotrophic factors (PDGF, NT-3 and IGF-2). The transdifferentiation of MDSCs was initially induced by Schwann cell (SC) conditioned medium. MDSCs adopted a spindle-like morphology similar to SCs after the transdifferentiation. Immunocytochemistry and immunoblot showed clearly that the SC markers S100, GFAP and p75 were expressed highly only after the transdifferentiation. Flow cytometry assay showed that the portion of S100 expressed cells was more than 60 percent and over one fourth of the transdifferentiated cells expressed all the three SC markers, indicating an efficient transdifferentiation. We then tested neurotrophic factors in the conditioned medium and found it was PDGF, NT-3 and IGF-2 in combination that conducted the transdifferentiation. Our findings demonstrate that it is possible to use specific neurotrophic factors to transdifferentiate MDSCs into Schwann cell-like cells, which might be therapeutically useful for clinical applications.

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PDGF, NT-3 and IGF-2 in Combination Induced Transdifferentiation of Muscle-Derived Stem Cells into Schwann Cell-Like Cells

NT-3 and IGF-2 in Combination Induced Transdifferentiation of Muscle-Derived Stem Cells into Schwann Cell-Like Cells. PLoS ONE 9(1): e73402. doi:10.1371/journal.pone.0073402 PDGF, NT-3 and IGF-2 in Combination Induced Transdifferentiation of Muscle-Derived Stem Cells into Schwann Cell-Like Cells Yi Tang 0 1 Hua He 0 1 Ning Cheng 0 1 Yanling Song 0 1 Weijin Ding 0 1 Yingfan Zhang 0 1 Wenhao Zhang 0 1 Jie Zhang 0 1 Heng Peng 0 1 Hua Jiang 0 1 Joao carlos Bettencourt de Medeiros Relvas, IBMC - Institute for Molecular and Cell Biology, Portugal 0 Ethics Statement All animal experiments were approved by the Administrative Committee of Experimental Animal Care and Use of Second Military Medical University (SMMU, Licence No. 2011023), and conformed to the National Institute of Health guidelines on the ethical use of animals 1 1 Department of Plastic Surgery, Changzheng Hospital, Second Military Medical University , Shanghai , China , 2 Department of Plastic Surgery, No. 411 Hospital of CPLA , Shanghai , China , 3 Department of Neurosurgery, Changzheng Hospital, Second Military Medical University , Shanghai , China , 4 Department of Transfusion, Changhai Hospital, Second Military Medical University , Shanghai , China , 5 Department of Hematology, XinHua Hospital, Affiliated to Shanghai Jiao Tong University (SJTU) School of Medicine , Shanghai , China , 6 Department of Mathematics, Hong Kong Baptist University , Kowloon , Hong Kong Muscle-derived stem cells (MDSCs) are multipotent stem cells with a remarkable long-term self-renewal and regeneration capacity. Here, we show that postnatal MDSCs could be transdifferentiated into Schwann cell-like cells upon the combined treatment of three neurotrophic factors (PDGF, NT-3 and IGF-2). The transdifferentiation of MDSCs was initially induced by Schwann cell (SC) conditioned medium. MDSCs adopted a spindle-like morphology similar to SCs after the transdifferentiation. Immunocytochemistry and immunoblot showed clearly that the SC markers S100, GFAP and p75 were expressed highly only after the transdifferentiation. Flow cytometry assay showed that the portion of S100 expressed cells was more than 60 percent and over one fourth of the transdifferentiated cells expressed all the three SC markers, indicating an efficient transdifferentiation. We then tested neurotrophic factors in the conditioned medium and found it was PDGF, NT-3 and IGF-2 in combination that conducted the transdifferentiation. Our findings demonstrate that it is possible to use specific neurotrophic factors to transdifferentiate MDSCs into Schwann cell-like cells, which might be therapeutically useful for clinical applications. - Funding: This work was supported by National Natural Science Foundation of China (Grant No. 81100950. Website http://www.nsfc.gov.cn/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. . These authors contributed equally to this work. Schwann cells (SCs) play a crucial role in peripheral nerve development and regeneration, and are thus an attractive therapeutic target in peripheral nerve injuries [13]. It is reported that cultured SCs could induced neuronal sprouting and regrowth in cell culture experiments and improve peripheral nerve regeneration in vivo [4,5]. SCs can be obtained from nerve biopsies for autologous transplantation and will not elicit an intense immune response. However, its difficult to culture sufficient numbers of autologous SCs because of their restricted mitotic activity, and there are also other disadvantages such as limitations in the supply of nerve material [6,7]. Use of allogeneic cells would need subsequent clinical immunosuppression [4]. Stem cells may be an alternative source for SCs. However, the clinical application of embryonic stem cells is limited because of ethical problems and their carcinogenic potential [8]. Increase evidence shows that adult stem cells may be promising candidate sources of cells [9,10]. Skeletal muscle may represent a convenient and valuable source of stem cells for stem cell-mediated gene therapy. Previous evidence supports the existence of MDSCs that exhibits both multipotentiality and self-renewal capabilities and therefore can be used for tissue engineering and regenerative therapy [11,12]. MDSCs have the ability to differentiate, upon stimulation with defined media, into multiple types of cells, including myogenic, hematopoietic, osteogenic, adipogenic, and chondrogenic-like cells [13]. The apparent advantages of MDSCs have led us to investigate whether they could be transdifferentiated to a Schwann cell phenotype. Our aim was to assess the phenotypic and bioassay characteristics of MDSCs transdifferentiated to SC-like cells. Importantly, we also sought to determine the neurotrophic factors which directed the transdifferentiation. Materials and Methods Isola (...truncated)


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Yi Tang, Hua He, Ning Cheng, Yanling Song, Weijin Ding, Yingfan Zhang, Wenhao Zhang, Jie Zhang, Heng Peng, Hua Jiang. PDGF, NT-3 and IGF-2 in Combination Induced Transdifferentiation of Muscle-Derived Stem Cells into Schwann Cell-Like Cells, PLOS ONE, 2014, Volume 9, Issue 1, DOI: 10.1371/journal.pone.0073402