3 years ago

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells.

Rui-Fang Li, Guo-Xin Nan, Dan Wang, Chang Gao, Juan Yang, Tong-Chuan He, Zhong-lin Zhang

Background : The effects of Simian virus 40 T antigen (SV40T) on various kinds of cells are different. Previous researchers failed to use SV40T immortalized nerve cells. However, they argued that SV40T caused nerve cell transformation. No one further study what is the specific effect of SV40T on nerve cells. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T in order to explore the effects of SV40T on Schwann cells.

Methods : SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, as well as co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry.

Results: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus . In addition, SV40T up-regulated the neurocrest markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the late differentiation markers MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells.

Conclusions: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells that can differentiate to terminal nerve cells.

Publisher URL: https://www.researchsquare.com/article/rs-17698/v1

DOI: 10.21203/rs.3.rs-17698/v1

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